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BATCH NUMBER 105587 ORDER NUMBER 91976 DESCRIPTION OF THE PROBLEM No signal. SAMPLE rat tissues; rat brain primary culture DETECTION METHOD ECL, ECLPlus ANTIBODY STORAGE CONDITIONS -20?C AMOUNT OF PROTEIN LOADED 50 ug ELECTROPHORESIS/GEL CONDITIONS 10% HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 3 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes
Asked on Sep 30 2005
Thank you for your enquiry. ab5295 is currently a fast-track antibody and has not been fully characterized. According to the datasheet there is a 76% identity between the immunogen and the corresponding rat homolog. I would therefore consider it highly possible that the result you are obtaining is a result of a failure of the antiserum to cross react against the rat RanBP9. However, there are several steps and controls that you could include to fully determine whether this is indeed the case. 1. Include a positive human (e.g. HeLa) cell lysate control (~20ug). 2. Try to use PVDF membrane. Generally, we find enhancement of transfer of proteins on PVDF membranes compared to nitroCellulose membranes. 3. Dilute the antibody 1:500 and incubate the filter overnight at 4 oC to see whether you are obtaining any signal whatsoever. 4. If you have enough material, try to load at least 20-30 ug protein/lane. Please let us know how you got on using this antibody by leaving an Abreview and in return we will award you 50 Abcam Points, which can be redeemed on a number of rewards (a further 100 Abcam Points will be offered for an image). Should you be the 1st researcher to leave positive or negative (conclusive) data in an Abreview for this product we will award you 1,000 Abcam points.
Answered on Oct 03 2005