Question (18113) | Anti-FSBA antibody [2F5/5] (ab18575)

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Question

BATCH NUMBER 178694 ORDER NUMBER HC41681 DESCRIPTION OF THE PROBLEM Does not recognise anything. SAMPLE HeLa lysate. PRIMARY ANTIBODY FSBA antibody, Abcam, 1h @ 1:2500 DETECTION METHOD ECL POSITIVE AND NEGATIVE CONTROLS USED Samples with no FSBA added. ANTIBODY STORAGE CONDITIONS Aliquoted and stored at -20. SAMPLE PREPARATION Basic lysis buffer with protease inhibitors stored at -80. AMOUNT OF PROTEIN LOADED 100 mcg. ELECTROPHORESIS/GEL CONDITIONS reducing, 10% TRANSFER AND BLOCKING CONDITIONS powdered milk at blocking agent, Tris Glycine transfer buffer. SECONDARY ANTIBODY [another company] goat anti-mouse 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

Answer

Thank you for your enquiry. This antibody has not been previously tested in Western blot. Thus, we cannot guarantee its suitability in this application. However, I would recommend trying this at a lower dilution, such as 1/1000. Since this is provided at a concentration of 0.9mg/ml, you are using less than 0.5ug/ml of antibody, which is rather low. I generally recommend using a few different concentrations between 0.5ug/ml and 5 ug/ml. I would also recommend incubating overnight at 4C. You did not mention your blocking conditions. I would recommend blocking with 5% milk in TBST for 1 h at room temperature, and incubating with the primary in TBST alone with no milk. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

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