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I wonder if you could give me some advice please.
I want to purchase an antibody which can be used to detect ALL 9isoforms ofPDE4D (phosphodiesterase type 4D). From what I can gather, you have 16 antibodies for PDE4D, I have purchased one previously (ab14613) but this is only specific toisoforms PDE4D1, PDE4D2, PDE4D3,PDE4D4 and PDE4D5.
The only other which appears to be similar to what I'm looking for is ab59249, whichstates "This antibody detects endogenous levels of totalPDE4D protein". However, when looking further down the page at the images, there appears to be only one band in the resulting WB. This is strange since that of the 9 isoforms, 6 are long and 3 are short i.e. I would expect to see 2 bands on the WB (as you do with ab14613).
Please could you explain the situation and also advise on which antibody I would be best purchasing?
Asked on Aug 23 2012
Thank you for contacting us. I apologise for the delay, I wanted to check this case with my lab colleagues.
Regarding ab59249, the amino acid range of this immunogen is aaXXXX of human PDE4D with accession# Q08499 (http://www.uniprot.org/uniprot/Q08499).
This antibody binds to isoforms 4, 3, 5, N3, 8, 9, 7, and 12. It does not bind to isoform 10, 1, 2, and 6. According to the isoforms that this antibody binds, you would expect to see two bands, approximately 25 and 85 kDa. Because the western blot on our website is cut off at 34 kDa, we can retest this antibody to see if it does actually bind the lower isoform as well.
Although we haven’t specifically tested the antibody against all isoforms, there are other alternatives that may also recognise several forms of the protein:
For ab77080 we ran a BLAST search and found the antigen sequence in all of 9 isoforms:
Ab112018 also shares 100% identity to isoforms PDE4D1-9:
Ab99409 and ab87329 recognize 10 of 12 isoforms listed for SwissProt entry Q08499. The two isoforms listed for SwissProt Q08499 not recognized by either of them are sp|Q08499-7|PDE4D_HUMAN and sp|Q08499-12|PDE4D_HUMAN:
And ab87329’s immunogen is present in 8 out of 12 of them so it should detect isoforms 1- 10.
I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.
Answered on Aug 23 2012