Talk:Ligand binding assay

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Have expanded article a bit, but it obviously needs much expansion, perhaps including a Scatchard plot and/or a standard curve graph etc.FeatherPluma (talk) 07:07, 5 September 2011 (UTC)

This topic is being edited as an assignment in an undergraduate neurobiology course. The course is participating in the Wikipedia Education Program. The revised article will be posted by March 24, 2014. Sargento21 (talk) 00:35, 15 February 2014 (UTC)

PLEASE NOTE: Ligand Binding Assay is a broad category for which many sub-analyses are utilized. Being said, there is an overwhelming amount of information for an assignment of this magnitude and not every type of Ligand Binding Assay is noted below. Thus, the following information indicates the most prominent methods in current research, as well as its applicability to future directions in the field. Keeping this in mind, however, all comments and suggestions are welcome. — Preceding unsigned comment added by MU Senior2014 (talkcontribs) 18:35, 1 April 2014 (UTC)

Primary Review #1[edit]

Overall this article was very well written and thorough for what you chose to cover on your topic. I liked how the flow was very homogenous between sections as well. You did a good job collaborating and making the article feel like it had one author. I have a few specific suggestions that I think would improve your page. In the Applications section, you refer to a different section in your own article. You can actually link to this section in your article, which I think would look better than just saying “see radioactive section”. Although I will say about this suggestion that the coding to accomplish this is complicated and I’m still trying to figure it out in my own article, so that’s not a very pressing issue. I would suggest that you split the section that includes radioactive and non-radioactive assays. I think it would look better and be a bit clearer if the broad categories were “radioactive” and “non-radioactive” rather than these being smaller categories in a larger one that simply combines them. It seems superfluous to organize it this way. In the radioactive section before you give specific radioactive assays, I think it would be useful to link a few more technical terms, like “specific activity” for example. Also, there a few places in your article where your category heading matches a link in that section, specifically in “fluorescence resonance energy transfer” and “surface plasmon resonance”. The way a lot of articles handle this is to include a link right below the heading that says “main article: X”. I think it would be useful for you to do this as well. The information in those sections is good, but it would be nice for clarity to make it obvious that there is an entire article on your broad section. I think you should do the same “main article: X” formatting with “real time polymerase chain reaction” as well as including a sentence or two of background info within your article. I’m sure you’re aware of this, but just to be thorough I’ll mention that there is no information under two of the solid phase ligand binding assays. As for an illustration, I think it would definitely be useful, but I’m sure you ran into the problem of finding one to use that is in the public domain. We’ve all seen hundreds of images in textbooks that would be perfect to use to show ligand binding assays, but you can’t use any of these copyrighted materials. So while it would be useful to include a diagram of at least one of the specific assays, I understand that it might not be feasible. I did think the graphics you included were both valuable and sufficient for this article.

Your sources are abundant and contain no original research. I liked how you had external links to many of them. I chose to review your fourteenth reference, which was a review in the British Journal of Pharmacology. This was definitely a secondary medical source because it was a review published in a medical journal. The information you cited with it was accurate, however in the “filter assays” section you cite it many more times than you need to. Site in once at the end of the information rather than after each successive sentence.

JBMarquette (talk) — Preceding undated comment added 13:42, 3 April 2014 (UTC)

Response to Primary Review #1[edit]

JBMarquette Thank you for reviewing our article and for your input. Here are the changes we have made based on your thoughtful suggestions, and if we did not decide to make changes the explanations as to why:

As you suggested, we linked the “see radioactive below” so that the user can just click on the word and jump straight to what we were referring them to. Next, we split up the Radioactive and Non-radioactive section. You were correct, it looks better, and it is more clear to identify the categories. We chose not to link Fluorescence resonance energy transfer or surface plasmon resonance directly under the heading because we already have them linked in the paragraphs below. For us it just did not seem necessary to link them directly below again. We attempted to like additional terms in the radioactive section like you suggested where applicable. --Jordannetts (talk) 02:54, 24 April 2014 (UTC)


Primary Review #2[edit]

This is a well written article with immense amounts of information. Considering the plethora of resources and wide range of usage, I think this article does a good job of distinguishing various types of LBA's based on characteristic aspects of the assay, such as the presence of radioactive elements or the physical phase of the material (liquid vs solid). My biggest critique is that at times the article is not reader friendly, especially to a non-scientifically literate person.

  • Introduction: The introduction does a nice job referencing what key terms are important, such as an assay or ligand/receptor. It is important to familiarize the reader with the material and help contextualize the topic, which I think your introduction achieves. The intro could use some examples of why ligand binding assays are so important, such as being used as a diagnostic tool or to help map out a biochemical pathway by detecting the presence of reactant/intermediate/product. It would also help giving some early examples of a ligand binding assay, such as ELISA, certain chromatography techniques, and southern blotting. Bringing up a concept such as enzyme kinetics may also be helpful in the intro. The rate of ligand "binding vs release" and how long the ligand stays bound to the receptor are always vital calculations to any ligand binding assay and may help get the reader get acquainted with the calculations and concepts addressing the affinity of ligand binding later in the article.
  • History: This section needs to be more reader friendly. It is good to address the reason for LBA's need and importance, and how/why it became such a common laboratory technique. That being said all of the details about the initial discovery/use are unnecessary. Simply address why it is was needed and how it became discovered/used for the first time, and why it became such a widely used technique.
  • Application of LBA: There is an overabundance of applications of ligand binding assay's so this is a difficult section and may be impossible to provide a comprehensive description. You do a nice of job of summarizing a typical ligand binding assay such as ELISA, the different affinity rates one might measure, and essential variables to the assay such as molarity. Also the last sentence is very good, showing some limitations to the LBA. Since the title of this section is application, I thought you could add some clinical or research applications, such as diagnostic validity scores.

There are various ways to distinguish different types of ligand binding assays. I thought your chosen distinctions between radioactive vs non-radioactive, and solid phase vs liquid phase did a fine job of classifying and organizing various LBA's.

  • Radioactive vs non-radioactive LBA's: The radioactive section containing topics such as saturation binding, the scatchard plot, competition binding were all informative and properly classified. The nonradioactive section provided a very nice assortment of commonly used techniques such as FP and FRET. The descriptions of these techniques were accurate and demonstrated why each technique had value and what it accomplished.
  • Liquid vs solid phase LBA's: The liquid phase section is good and provides numerous examples of very common techniques such as ELISA and western blotting. I know that ELISA uses a multiwell plate design so I was confused why multiwell plate designs were specifically labeled as only a solid phase assay. Both sections present various assays that involve ligand binding and accurately describe what the assays measure and how quantitative analysis is achieved.
  • Selectivity and specificity: This is a major concept in any type of chemical binding, the distinction between how selective the ligand is and how tightly does the ligand bind to its target. This section can do a better job of distinguishing these two concepts and give examples of various ligands and their relative affinity and specificity to various receptors.
  • Technological advances: Advances in technology are always critical to the advancement of any assay. The section does a nice job highlighting improvements in techniques and technology that may be vital for future research.

I reviewed the following reference: Joseph R. Lakowicz. (1991) Topics in Fluorescence Spectroscopy: Biochemical applications. This was an appropriate secondary source which discussed numerous ligand binding assays, which isolated or detected a variety of different macromolecules. This reference along with the other references are all appropriately cited throughout the article.

Overall, the article was

1) Well written and contained a large amount of information about a very broad topic. 2) Verifiable with lots of appropriate sources 3) Neutral and fair. 4) Illustrated with nice figures and sample equations. Some pictures of ligand molecules, receptors, or pictures of LBA experimental setups would be visually pleasing.

MU77 (talk) 18:21, 6 April 2014 (UTC)

Response to Primary Review #2[edit]

MU77, thank you for your response and thoughtful input in writing a review for ligand binding assay. Here are the changes we made by section and if relevant, explanations for why we did not make your suggested changes.

Introduction:

  • Although you had many good suggestions for the introduction, we thought adding any more might make it too long. We felt that our lead paragraph summarized the subject according to Wikipedia standards because it gives the main ideas without going into too much detail.

History:

  • The details about the initial discovery in the second paragraph were present on the stub before we began this project. We found it to be repeated in a few history sources about LBA and decided it was interesting and necessary to include/keep in the article.

Application:

  • Thank your for the complements on this section. Other users suggested research applications as well and we found that the sources with this information were not secondary sources, therefore, we did not want to interfere with the neutrality of Wikipedia articles given the currently available sources on LBA.

Radioactive vs non-radioactive:

  • We have changed the flow of these sections to be more distinctive by categories, therefore, making it easier for the reader to flow through the article.

Liquid vs solid phase LBA:

  • Again thank you for your complements. As far as separating ELISA and multiplate assay, we felt that there is a important difference and a large amount of information on both that multiplate and ELISA both needed separate sections.

Selectivity and specificity:

  • We have changed this section to ‘Binding Specificity’ as well as making this section’s significance more clear to the reader. This made the section as a whole easier to comprehend. We included a more detailed explanation for what makes a binding relationship specific or nonspecific.

Technological Advances:

  • Thank you for your comments on this section.

Overall:

  • Since your review, we have added multiple images to various sections of our page that you might now find more visually pleasing. Sargento21 (talk) 03:46, 24 April 2014 (UTC)


Primary Review #3[edit]

By section:

1) Introduction- The author has a clear introduction to the topic. Although the method is prefaced in this section, some basic introduction to examples of its use in industry would be helpful. Where is this used in the field? When did it it become a prominent assay (if it did at all)? I know the following section is history, but it is helpful when the introduction gives a very general idea of the what it is, when it first came into practice, and what is it used for today. That way, the reader gets a general idea of the assay, and if the reader wants more information, they can refer to the following sections.

2) History- The author explains well what the assay was used for in the past, and the transition into immunohistochemistry. Citations are needed for the first two sentences of this section. Maybe some information to include is: who is accredited with discovering this method? Was this used in any famous, historical studies? When was this assay created and when did IHC become more prominent? Can any photos of the procedure be added? Please check this section for grammar. Some articles are missing, and there are some sentences that can be more concise.

3) Application Ligand Binding Assays- This brief paragraph did a great job of providing neutral information by providing benefits and drawbacks to the procedure. Could some information about the methods be provided? Is there any general knowledge about the the assays, such as relative expenses or ease with which the procedure is carried out?

4) Radioactive and Non-Radioactive Ligand Binding Assays- For the saturation binding, where is saturation binding used? What are some clinical and/or other practical applications of this method? It would be helpful if this section included a brief methods section for the saturation binding, and the competition binding experiments. For the sake of clarity, the section would flow better if all of the radioactive assay descriptions were following each other (such as saturation binding and competition binding). The non-radioactive section was clear with information, grammar and punctuation needs to be reviewed.

5) Liquid Phase Ligand Binding Assays- Clear and concise!

6) Solid Phase Ligand Binding Assays- Multiwell Plate Assays: are these manufactured or made by investigators themselves? Can a brief instruction of how to use them be incorporated in this section? If a picture could be inserted for this section, that would be helpful! It appears that two sections are missing content.

7) Selectivity and Specificity- What characteristics of a ligand and its respective receptor make them selective or specific?

8) Limitations- Great section, some pictures would improve it!

Source reviewed: Cereto-Massagué, Adrià; María José Ojeda, Robbie P. Joosten, Cristina Valls, Miquel Mulero, M. Josepa Salvado, Anna Arola-Arnal, Lluís Arola, Santiago Garcia-Vallvé, and Gerard Pujadas. (201). "The good, the bad and the dubious: VHELIBS, a validation helper for ligands and binding sites". Journal of Cheminformatics 5 (36): 1–9. doi:10.1186/1758-2946-5-36.

  • This article does not appear to be a secondary article. It seems to be a primary source, seeing that the authors provided their own data in the results section. Wikipedia asks for secondary sources, such as reviews. Seeing that this source was used for relatively general information, the same information should be found in a secondary source. The information cited by this source is all within the text. It is recommended, however, that the author find an alternative secondary source.

Overall, the article was very informative, well organized, and well cited. If possible, more methods and details about the respective techniques would be helpful, as well as some pictures or diagrams to illustrate the assays. The article should be reviewed for some grammar and punctuation. Ideas seem to be broken up in parts due to the punctuation errors.

NWcoffee (talk) 06:33, 7 April 2014 (UTC)

Response to Primary Review #3[edit]

NWcoffee, Thank you very much for your thorough and well-written review. Your comments, suggestions, and overall feedback were very helpful for our page and we appreciate your help. That being said, I would like to share with you what we have implemented, changed, and taken away per your suggestions. Just as your review, I will present the following information by each respective section.

Introduction:

  • Your suggestions to provide basic examples of the use of ligand binding assay, as well as when LBA became prominent in the industry was very helpful. Being said, we have added information regarding its use in the field at the cellular level, as well as the broader advances within pharmacology. We agreed with your suggestion to include information about the prominence of LBA within the field; however, instead of providing this information in the introduction, we have included such information within the history section, as the information seemed to suit this section better than in the introduction. Nevertheless, your suggestion was very helpful for us to include more background information.

History:

  • Your feedback in regards to this section was incredibly helpful. As stated above, we have combined your feedback about ‘prominence within the field’ with our ‘history’ section. Being said, we added relevant information pertaining the accreditation and historical studies. In regards to your question about when immunohistochemistry became more prominent in investigating estrogen receptors of various breast cancers, it is inferred that this technique became relevant around 1998-1999. However, after further investigation of this information, it appears that this specific section is reported from a primary, versus secondary, article. Being said, we have taken out this information due to the parameters of citations within Wikipedia. Nevertheless, thank you for pointing out this hole in information, as it has made us reevaluate the sources drawn upon.
  • In regards to images for the procedure of LBA, we have added many more images throughout the rest of our page of the various types of LBAs!
  • Lastly, we appreciate your feedback regarding grammar, as there were quite a few errors.

Applications for Ligand Binding

  • Due to the parameters of this assignment, we have avoided providing any methods about how to do any LBAs. It was a great suggestion, however.
  • General knowledge about the assays regarding relative expenses depends on the specific assay, which opens a plethora of options, such as which company to buy from, the desired quantity, etc. However, this information has not been found in secondary sources and will be left out as the result.
  • Your suggestion regarding the ease with which the various procedures are carried out is helpful, but we. The ‘ease’ depends on type of assay researchers are using, techniques used, researcher experience, etc. However, because we provide the benefits and uniqueness of each type of LBA throughout our page, we feel that it would be redundant to incorporate the ease of each technique in this section.

Radioactive and Non-Radioactive Ligand Binding Assays

  • In regards to your question, “where is saturation binding being used”, we have added this information, as it was surprising that we did not include it in the first place. Thank you for pointing this out.
  • As helpful as your question/feedback, “What are some clinical and/or other practical applications of this method? It would be helpful if this section included a brief methods section for the saturation binding, and the competition binding experiments” is, it goes beyond the magnitude of this assignment.
  • You were not alone in providing very helpful feedback regarding these sections – they needed some polishing. Being said, we have changed how we split these sections by making them 2 distinct sections, versus 2 subsections under the heading “Radioactive and Non-Radioactive Ligand Binding Assays”.
  • Thank you, once again, for your reminder to review grammar. Thus, grammar, punctuation and spelling were edited.

Solid Phase Ligand Binding Assays

  • Your question regarding the manufacturing or multiwell plate assays was incredibly helpful, and a great question at that. Being said, the multiwell plates are manufactured and the assays that are tested within the plates are influenced by the researchers themselves. We have added not only clarification about this, but also an image that displays what the multiwell plates look like in order to decrease confusion. Thank you for pointing out this confusion!
  • Instead of incorporating the step-by-step instruction on how to use/run multiwall assays, we have added more in-depth information about the information obtained from such assays.
  • Since your request for an image, we have found a great picture of various types of multiwell plates and have added it within this section.
  • We are aware of the two empty sections and have been debating whether or not to keep them, as we had a difficult time finding secondary sources to support these types of assays. In addition, there was a lack of explaining these techniques and due to the nature of this assignment, we decided to acknowledge its existence in the field, but leave it be. We hope that future visitors to our page can find further information to help build these two sections.

Selectivity and Specificity

  • Your question regarding the “characteristics of a ligand and its respective receptor” and its selectivity/specificity made us realize that this section is quite confusing. Thus, we have modified the subheading to read “Binding Specificity”, as it is known in the field, in addition to incorporating a more detailed explanation regarding what makes a binding relationship specific or nonspecific.

Limitations

  • Your suggestion to add pictures was fulfilled, as we added two images of what the PET scan machine looks like, as well as the type of information obtained after a PET scan has been fun.

Source Reviewed

  • Your article review was incredibly helpful, and we were rather dubfounded that we did not catch this mistake earlier. So, thank you for pointing this out, as we have deleted all of the information that was originally cited from this article.

Overall feedback/comments

  • All of your feedback, suggestions, comments, and overall support have been very helpful in the review process of this page. On behalf of our group, thank you for providing such detailed and thorough information! MU Senior2014 (talk) 01:50, 20 April 2014 (UTC)

Primary Review #4[edit]

Also by section:

1) Introduction:

  • Second sentence- replace “some kind of” with a different word or phrase

2) History:

  • The example provided in this section is very specific to a certain area of research. It might be helpful to provide a broader example or several specific examples from diverse fields of study. This section would also be rather difficult to get through for a lay reader.

3) Applications:

  • This section is a really excellent outline for what could be included here. I recommend expanding on the current content by providing clinical examples and the significance of this assay in research and clinical settings. Maybe discuss the advantages/disadvantages of this method vs. immunohistochemistry etc.

4) Radioactive

  • you could begin this portion of the article with a sentence or two that explains the radioactive method in the vernacular. This will help the uninformed reader understand the specific information you provide.

5) Non-radioactive

  • specify what the subheadings refer to...specific methods, how to label etc... otherwise the reader has to try and figure out what these subheadings are trying to tell them

6) Liquid Phase Ligand Binding Assays

  • use the entire term before providing the acronym (ie- ELISA) in this section. Otherwise this section was both quite informative and exceptionally easy to read and understand.

7) Solid Phase LBA

  • Remember to maintain the voice of an encyclopedia in this section. At some points it strayed into the territory of making suggestions on the proper methodology (ie- It is recommended)
  • This may be a good place to include an image of a well plate?

8) Selectivity and Specificity

  • If you are going to discuss selectivity and specificity provide super brief definitions of each of these words for the reader.
  • It may have just been me but I felt a bit lost on the significance of this section. It may be helpful to put the cited pieces of information in context by at least one additional sentence to explain each cited example.

9) Technological Advances

  • A brief sentence at the beginning of this section may help the reader transition into this portion of the article.

10) Source I reviewed “Immunoprecipitation (IP) technical guide and protocols” ThermoFisher Scientific Inc. I thought this was an excellent secondary source to use to provide reliable information about immunoprecipitation. I believe your use of this source was sufficient as it discussed only a very specific portion of your article. Nice find!

11) General remarks-

  • Possibly provide examples of situations in which each type of LBA would be used. It helps to place the methodology in context of real-world research and diagnostics.
  • You did a great job discussing the methods of a rather complicated measurement tool.
  • A very informative article. Most of my suggestions are just to improve readability; the content seems to be there. I thought you guys did a great job organizing a broad field into a few sections for the reader. Hopefully some of my suggestions will be helpful! MUhelb (talk) 12:12, 7 April 2014 (UTC)

Response to Primary Review #4[edit]

MUhelb, thank you for your response and thoughtful input in writing a review for ligand binding assay. Here are the changes we made by section and if relevant, explanations for why we did not make your suggested changes.

Introduction:

  • We took your advice on the introduction phrasing in that sentence and changed it to sound better.

History:

  • For the history section example, it was present on the LBA stub before working on the article. Although it is a very specific example, it is the research for which the foundations of LBA were built upon, therefore, we decided this example was relevant and interesting to keep in the article.

Application:

  • We appreciate these suggestions as well as the general comment you made related to this, but we believe that providing clinical examples and explaining the comparison against other methods would interfere with the neutrality of Wikipedia articles given the currently available sources on LBA.

Radioactive:

  • We took your advice on a general introductory sentence/transition statement at the beginning of the radioactive section to help the reader better understand what is going to be discussed.

Nonradioactive:

  • We have re-organized the radioactive and nonradioactive headings so that the reader can better navigate through these sections, so they are no longer getting lost in the subheadings.
  • We have added a few images to this section as well.

Liquid and Solid Binding Assay:

  • The ELISA acronym is now spelled out, and we have added a picture of a well plate.
  • We also fixed the paragraphs which strayed to talking about making suggestions on the proper methodology.

Selectivity and Specificity:

  • We have changed this section to ‘Binding Specficity’.
  • We made this section’s significance more clear and easier to comprehend by the reader.

Technological Advances:

  • This section now includes a brief transition sentence to help the reader better understand and transition into this part of the article.

General Remarks:

  • Thank you for the review of our source.
  • Using examples of the situations in which LBA would be used is a good suggestion, and we will go back and include some if the information is available in secondary sources if they do not interfere with the Wikipedia neutrality. Sargento21 (talk) 14:40, 21 April 2014 (UTC)

Secondary Review #1[edit]

I think you did a really good job on this article. There is a lot of information available, but you kept everything organized and explained things very well. The introduction was great because even though there is a lot of information to cover, you stayed as broad as possible (while providing meaningful information) so that people could find more specific information in the rest of the article. The amount of subsections you have and the way you organized them is probably the most impressive part about this Wikipedia article. It was a great idea to provide a general description of the type of assay, then link to a more specific article. The pictures of the equation, scatchard plot, and the table in the Selectivity and Specificty section are creative ways to make what you're talking about easier to understand. On the surface it appears like you could add some more pictures, but the pictures for the assays are generally already in the linked to articles. I don't think I have anything bad to say about this article, aside from a couple of empty subsections (which I'm sure you're getting to eventually), this is a well written article that is organized really well. Great job!--D23sunn (talk) 19:28, 30 March 2014 (UTC)

Response to Secondary Review #1[edit]

D23sunn, thank you for your response and thoughtful input in writing a review for ligand binding assay. We have added more images since your review to make the article more visually appealing. As far as the empty subsections, we felt that there was a large amount of information on the topic and attempted to narrow it down, but we added some additional subsections we found important so that in the future someone can fill that in once and when the proper sources become available. Thank you for your comments and suggestions. Sargento21 (talk) 03:55, 24 April 2014 (UTC)

Secondary Review #2[edit]

The page starts off simple and easy to understand, slowly working into the topic, which is really captivating. I like the equation added, not only does it add more verifiability, but further information for readers to apply even. Overall, the reading is very straight-forward and detailed, which I really appreciate over embellished work. I think if you read the article aloud, some of the wording could be changed for easier reading, but that is completely dependable on preference. Lastly, a few more images could add flavor to the article and is always helpful for visual learners to reach a broader audience. Concluding, the article was very well written and planned, I agree the organization is very well done. Great Job!! MissKell (talk) 08:40, 2 April 2014 (UTC)

Response to Secondary Review #2[edit]

MissKell, thank you for your review on ligand binding assay and your suggestions. Since your review, we have added more images to make the article more visually appealing. Thank you for the overall complements and on the organization of the article. Sargento21 (talk) 04:00, 24 April 2014 (UTC)

Secondary Review #3[edit]

The article starts off easy but becomes more dry the further you read. The equation was helpful but more images would make the article more visually pleasing. They don't have to be directly about LBA but can something simple like how antibodies bind to receptors (the kind of graphics we find in textbooks. I like how the article contains a lot of information and it is organized very well. It was hard to tell where the author of each section changed which made the article more seamless and complete. All in all a good article but add some color.Stockton Whitfield (talk) 13:53, 4 April 2014 (UTC)

Response to Secondary Review #3[edit]

Stockton Whitfield, thank your for your review of ligand binding assay. We have added more images to the article, which made it more visually pleasing. Thank your for the complements on the organization and information. Sargento21 (talk) 04:04, 24 April 2014 (UTC)

Secondary Review #4[edit]

This article is very long and presents a whole lot of information. That being said I think it is very important that the authors were able to make a working and easy to use content box as well as an article that flows naturally, rather than one that might seem out of order or incoherent at times. I do think that the authors will benefit with providing more images to break up some of the longer text sections but other than that I think it was a very well written and organized article that covered a broad range of this topic. --Tzanon21 (talk) 20:15, 6 April 2014 (UTC)

Response to Secondary Review #4[edit]

Tzanon21, thank you for your review on ligand binding assay. We appreciate your comments and suggestions. Since your review, we have added multiple images to the page to make it more visually appealing. Sargento21 (talk) 04:07, 24 April 2014 (UTC)

Secondary Review #5[edit]

I can tell a lot of time was put into this article because of the abundance of information. I thought the addition of the plot diagram, equation, and chart were very helpful and wise when trying to explain information. I only have a few suggestions/questions. It the history section, I believe there a needs to be a capital "I" on the word It in the second paprgraph. In the Saturation Binding section, if the sectences "there are two common strategies that are adopted for this type of experiment:[15] Increasing the amount of radioligand added while maintaining a constant specific activity.[15] Maintaining a constant concentration of radioligand and decreasing the specific activity of the radioligand (by addition of unlabeled ligand).[15]" was combined into one with "and" it would read better and so the readers knows those are the two types. Under the FRET section, I think the last sentence may sound better if the word "when" was added so the sentence reads "It is critical when using FRET..." Lastly, I was just curious why some headings had no information under them such as On-bead ligand binding assay and on-column ligand binding assay. Overall, I think the article was nicely done!Kkakes728 (talk) 00:35, 7 April 2014 (UTC)

Response to Secondary Review #5[edit]

Kkakes728, thank you for your thoughtful comments and suggestions on the article. We have changed the article based on your multiple grammar and sentence structure suggestions. The headings without any information under them were put there because we felt that they were necessary in the future direction of this article, but were out of the scope of this project just based on how long the article was already and the available information on the empty subheadings. Sargento21 (talk) 04:11, 24 April 2014 (UTC)

Secondary Review #6[edit]

It is obvious that this article was well researched and put together. I agree with reviewer #4 above that some images of the assays described would go a long ways to providing context to the large amount of text. Specifically, if you could find some real life pictures of the assays in a laboratory setting, instead of a cartoon, it may give context on how these assays are carried out in real life in a visual way. SamSchultz1 (talk) 01:28, 7 April 2014 (UTC)

Response to Secondary Review #6[edit]

SamSchultz1, thank you for your review on ligand binding assay. We have added images throughout the article, which makes it more visually appealing to the reader than it was prior, although we could not find real life pictures like you suggested. Sargento21 (talk) 04:15, 24 April 2014 (UTC)

Secondary Review #7[edit]

Well done! You guys did a great job. The article flows nicely and I can tell that you spent a lot of time on it. Being a visual learner, I liked how you had a visual aid to display the equation for the actual concentration of a radioligand and the scatchard plot. That being said, I think it would be beneficial for you to include a large photo at the beginning of the article, and Sam made some good suggestions in his secondary review above for that. Overall, great job! MKClement (talk) 20:26, 7 April 2014 (UTC)MKClement

Response to Secondary Review #7[edit]

MKClement, thank you for your comments and suggestions on ligand binding assay. Since your review, we have added multiple images throughout the article that we found makes it more visually appealing. We could not find a large photo to put at the beginning of the article that fit perfectly with the information, but that is a very good suggestion and idea if one ever become available.Sargento21 (talk) 04:18, 24 April 2014 (UTC)

Secondary Review #8[edit]

Great job! I thought the introductory paragraph really summed up the article nicely and gave the reader a good understanding of the topic. However, more interested readers could go on to pursue the more detailed and advanced material. I really liked the use of the formula and the table. Another image could be beneficial. The article was easy to follow and very well organized. EmmaAWeber (talk) 02:27, 8 April 2014 (UTC)

Response to Secondary Review #8[edit]

EmmaAWeber, thank you for your review of ligand binding assay. Since your review, we have added more images to the article, which makes it more visually appealing to the reader. Thank your for your complements on the article. Sargento21 (talk) 04:21, 24 April 2014 (UTC)