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I would really like to try your Fast Track Gli3 antibody (Ab6050). I have a chromatin IP assay with a FLAG tagged endogenous Gli1 protein that works very well in mouse. Since Gli3 is known to bind several identical targets to Gli1, I have a built in positive control for the antibody (Ptch1, Gli1, both of which give me 20-fold enrichment by qPCR). Unfortunately, I can not afford to buy your antibody without knowing if it works - indeed, since there is a 1 amino acid deviation from the antigen in mouse and a negative band in the Western blot there seems to be some doubt if it will work in mouse at all. I was wondering if you would consider giving me a small sample (I would only need 20ug) to try on my Chromatin IP? If I did get it to work, I would certainly buy more of it from you. If it does not work, it will be a controlled negative result, which might itself be useful information to you.
Asked on Oct 07 2005
Thank you for your enquiry. ab6050 is a fast track antiserum which is an antibody that is yet to be fully characterized and sold at a reduced rate of £ 125.00 for 100 µg. I would be very willing to provide you with the antiserum at a reduced rate (20% off) so that we may determine its success by chromatin immunoprecipitation. It sounds like you have a nicely controlled system which would be great to exploit using this antibody. So far we have not been able to detect the endogenous protein using human or murine cell lysates (please refer to the datasheet and the western blot Abreview left by Tian Yu). We believe this may be due to the low level of expression in these lysates. However, given that you have a binding site and a positive control for ChIP I would be very interested in your data. Can I ask whether you have a FLAG tagged construct of Gli3, such that you would be able to directly compare the efficiency of ab6050 using ChIP with that of your FLAG tagged antibody under ChIP conditions. Please get back to me if you are interested in my offer. I look forward to hearing from you.
Answered on Oct 10 2005