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BATCH NUMBER 124239 ORDER NUMBER 92612 DESCRIPTION OF THE PROBLEM No signal or weak signal SAMPLE Smooth muscle cells (human and mouse), Mouse aorta tissue extract DETECTION METHOD Pierce SuperSignal West Piko Chemiluminescent Substrate POSITIVE AND NEGATIVE CONTROLS USED Control for secondary antibodies - used primary anti human/mouse GAPDH ANTIBODY STORAGE CONDITIONS -80 C SAMPLE PREPARATION RIPA + protease inhibitor coctail AMOUNT OF PROTEIN LOADED 30 ug ELECTROPHORESIS/GEL CONDITIONS Reduced 8% gel TRANSFER AND BLOCKING CONDITIONS Tris/Glycin buffer + 10% Methanol Tris buffer + 5% Milk (no fat) SECONDARY ANTIBODY Pierce, anti mouse, 1:10000, 1:5000, 1:4000 1,5 hour Tris buffer + 5% Milk (no fat) HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 6 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes
Asked on Aug 09 2005
Thank you for your enquiry and I'm sorry to hear that you are experiencing difficulty with ab8976. Can you please provide me with a few additional details: What dilutions of ab8976 have you tried? Do you know that the secondary antibody is working properly? Also, did you ensure that the protein transferred properly to the membrane? Thank you again, and i look forward to hearing from you.
Answered on Aug 10 2005