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What were the Western blot conditions used for the blot on the datasheet?
Asked on Jun 06 2006
Thank you for your enquiry. The membrane used was 0.45micron nitrocellulose, blocked in 5% non fat milk in TBS, antibody incubations in TBS +0.1% tween 20, ab7255 conc was equivalent of 1:500 from our standard tissue culture supernatant prep and incubation time was 1hr at RT with shaking. Detection in this case was with Goat anti-mouse alkaline phosphatase conjugate and development was with BCIP/NBT. The blot was made quite a while ago, probably about 1993, and if we were doing it today we would use chemiluminesnce and probably 1:1,000 to 1:2,000 ab concentration. I hope this information helps. Please do not hesitate to contact us again if you need anything further.
Answered on Jun 07 2006