Question (17464) | Anti-KIF4A/KIF4 antibody (ab3815)

Go to datasheet (ab3815)

Question

BATCH NUMBER 185414 ORDER NUMBER -- NOT SPECIFIED -- DESCRIPTION OF THE PROBLEM weak signal compared with batch number 162899 SAMPLE whole cell lysate isolated from human cultured DMS114 cells and DMS273 cells PRIMARY ANTIBODY 1:200 for 1 hour at room temp. DETECTION METHOD HRP-based method POSITIVE AND NEGATIVE CONTROLS USED no ANTIBODY STORAGE CONDITIONS Cosmo received it on 21/April/2006 and stored it at -20 C. The customer received it on 24/April/2006 and has stored it at -20 C. SAMPLE PREPARATION RIPA buffer AMOUNT OF PROTEIN LOADED 7ug/lane ELECTROPHORESIS/GEL CONDITIONS 7.5% SDS page gel TRANSFER AND BLOCKING CONDITIONS blocking agent; BLOCKACE, for 1 hour at 20 degrees SECONDARY ANTIBODY anti goat IgG-HRP 1:20000, for 1 hour at room temp. ADDITIONAL NOTES The customer had purchased batch 162899 of this antibody and gotten a good result in this system. Do you have another batch in stock now? Please give us your suggestion about this matter. He thinks its reproducibility is too weak.

Answer

Thank you for your enquiry. I am sorry to hear that you have been having difficulties with this antibody. We do experience occasional batch variation. This often means that it may be necessary for our customers to perform an element of optimisation in their experiments when receiving the antibody. On this occasion I would like to recommend that your customer increases the mass of protein that they are loading on the gel as we typically recommend that 20-30ug of protein is loaded. I would also recommend that the antibody is applied at a dilution of 1:100; as detailed on our datasheet using an overnight incubation. Taken in combination this is very likely to improve the results that your customer has been obtaining significantly. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

Sign up