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Your product literature says the the immunogen used to raise this antibody was recombinant full length human TGFbeta1 expressed in CHO cells. I would like to know if this means the mature protein, or the precursor protein.
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ANSWER: |
The immunogen was the mature protein. Please contact us again if you have any additional questions.
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BATCH NUMBER 80204 ORDER NUMBER 346421 DESCRIPTION OF THE PROBLEM Non-specific bands SAMPLE human myocardium tissue PRIMARY ANTIBODY Anti-TGF beta1 in different dilutions: 1:200; 1:500, 1:1000 SECONDARY ANTIBODY HPR-Anti-chiken antibody, dilutions: 1:1000, 1:10000, 1:20000 DETECTION METHOD ECLPlus, ECL POSITIVE AND NEGATIVE CONTROLS USED Multimarker ANTIBODY STORAGE CONDITIONS plus 4C (9 days) SAMPLE PREPARATION SDS sample buffer, heating 10 min (70C) AMOUNT OF PROTEIN LOADED 50 ng ELECTROPHORESIS/GEL CONDITIONS Reducing 4-12% gel, MOPS buffer(Invitrogen) 1 hour TRANSFER AND BLOCKING CONDITIONS Transer buffer (Invitrogen), blocking of non-specific binding by 5% milk over night HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 4 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? None ADDITIONAL NOTES I have bands everywhere but only not in the specific area
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ANSWER: |
I'm sorry to hear you are having problem with ab10518. I would like to suggest the following modifications to your protocol: -make sure your protease inhibitors are fresh when added to the lysis buffer -Use loading buffer made up with betamercaptoethanol and SDS and boil at 95-100 degrees celcium for 5min -block in BSA 5% and incubate your antibodies in TBST -test the secondary antibody: without primary and with other primary antibodies as this can go off -wash between steps extensively in TBST Please let me know if this helps and do not hesitate to contact us for further advice,
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