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Hello, I have a few questions about your product Anti-Factor VIII antibody (ab61370), which is sheep polyclonal anti-human FVIII antibody. 1. I am looking for an antibody to do sandwich ELISA for hFVIII detection in plasma. Is it feasible to use this Ab as capture Ab, and conjugate it with HRP and used as detection Ab? 2. Since I'll detect hFVIII in mouse, rat, pig and dog blood, I wonder if it has cross activity with other species. 3. Could you provide a few reference papers about ELISA assay with this product? I'll appreciate if you could reply asap.
Asked on Jul 12 2012
Thank you for contacting Abcam.
I do not think that you would be able to use the same antibody as both the capture and detection antibody in your sandwich ELISA, as you would want each antibody to detect different portions of the protein.
Also, we have not tested the antibody on any other species than Human, we cannot guarantee that it will work or not for your other species of interest. We normally look for at least 85% level of homology between the immunogen sequence and the target sequence, before we fell comfortable recommending an antibody. Based on the sequence alignments I performed (see results below), there is a chance that it may work with dog, however the level of homology with the others is low.
Pig - 75%
Dog - 85%
Mouse - 82%
Rat - 66%
You could take advantage of our testing discount program, if you want to try the antibody in dog tissue, you can find more information on that program below:
I have attached the general ELISA protocol that was used to test this antibody, but we do not have any other information that I can provide. We do put all relevant information, including references on the webpages, if we know about them.
If there is anything else I can help you with, please let me know.
Answered on Jul 12 2012