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Thank you for your reply, and this is my protocol.
1. I transfected MDA cells in lane 2 and hTERT cells in lane 4 with the TNF-alpha gene. Lane 1:MDA-vector lane4:hTERT vector.
2. Lysed the cells with Ripa buffer.
3. 25 ug loading. DTT+loading dye.
4. Blocking by 5 % dry non fat milk with 1 % BSA in TBST.
5. Anti-TNF dil 1:1000. 4 C overnight.
6. Secondary Ab is 1:5000 anti mouse. 1 hr RT.
but I have no idea about PO number.
Asked on Aug 14 2012
Thank you for your reply. Based on the protocol you provided, I would not expect the bands observed on your blot to be specific. I was able to locate an order that was shipped to *** last week for ab1793 and ab15563. Is this the correct order? If I am able to locate your order, I would be happy to offer you a free of charge replacement, credit, or refund. Please let me know how you would like to proceed and I will be happy to help you further.
Answered on Aug 14 2012