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To answer your questions: 1. I have tested TGF-b, b-actin and AIF on the same membrane. They all works fine. But no other anti-angiopoietin-1 antibody was used, except Ab8451 and Ab95230. Attachment is the picture of blot with b-actin. 2. 50ug/lane protein was loaded to gel. 3. Yes, Laemmli Sample buffer was from Bio-rad (161-0737), prepared 950ul of sample buffer with 50ul b-ME. Then samples were boiled at 95 C for 5 min. 4. Yes, they are the same, cause I tested them on the same membrane, one after another with stripping, washing and re-blocking between two probes. Hope that will be helpful. Thank you very much!
Asked on Jan 04 2012
Thanks for the additional information. A couple more questions... 1. Are one or more of the samples from the blot considered a negative control, in that it is known to not express Angiopoietin 1? 2. After stripping but before reprobing, was it determined that there was no residual ECL signal remaining from the previous blot? Thanks again!
Answered on Jan 04 2012