Talk:Digoxigenin

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Untitled[edit]

It can easily be attached to oligonucleotides, at either 5' or 3' ends by chemical modifications.

To detect mRNA production in a given cell :

1- Prepare complimentary RNA with DIG attached to them. Nowadays, you order that over the internet.

2- Following your preferred protocol for In situ hybridization, attach this probe to mRNA produced by the cell.

3- Use an anti-DIG antibody to detect DIG (and hence, mRNA).

This is basically rubbish. I'm not sure how Dig is attached, but it's usually conjugated to a specific nucleotide (normally Uridine) which is incorporated into the complimentary mRNA when it's synthesised - therefore attaching it to the bits where nucleotides join together into strands is unlikely.

Nobody in their right mind would order RNA over the internet; everyone but everyone produces their mRNA fragments in house. Presumably whoever wrote this is getting confused with primer design, which is (as far as I'm aware) largely done online; given oligonucleotide primers (10-20 nucleotides) and a cDNA library or a complete set of genomic DNA, you can produce a large number of gene fragments (commonly 100-1000 nucleotides) using PCR; which is usually put in a plasmid. You then use an RNA polymerase, and a nucleotide mix containing only Dig-labelled Uridine triphosphate, to make a transcript that can be detected by antibody.

It can also be conjugated to specific sugars and allowed to incorporate into glycoproteins in live cells, and detected by anti-Dig antibodies.

Am updating to reflect this, and to make it read less like a protocol for ISH.

Confuseddave 20:03, 12 January 2007 (UTC)[reply]

Should the first line say antigenicity instead of immunogenicity? I thought that haptens only elicited immune responses when conjugated to larger moleculars

99.231.52.177 (talk) 05:06, 28 January 2009 (UTC)[reply]

PubChem says that Digoxigenin can be isolated from D. orientalis and D. lanata, making no mention of D. purpurea. ~~I don't know what the truth is here, but it would seem that, at the very least, "exclusively" isn't the right word to use here.~~ Looked it up in Biochemical targets of plant bioactive compounds by Gideon Maxwell Polya (it's on Google Books) and it turns out digoxigenin present in all 3.

Dewert (talk) 16:43, 21 July 2009 (UTC)[reply]