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Hi, I ran a western blot yesterday using antibodies against |BIRC3(cIAP2). Attached picture is in Gel format. I am pretty sure you will be able to open with any wester blot analysing software. I used the abcam monoclonal antibody on the left membrane and polyclonal that you sent me on the right membrane. antibody concentrations were 1/500 for both antibodies. I loaded HELA, Daudi , A431 in the same sequence as in here. I blocked with 5% BSA for 1h, then incubated with antibodies overnight. Then I used an HRP linked secondary in 1/1500 concentration (incubated for 1h). I followed the protocol for incubating with ECL then exposed at different time points using ImageQuant LAS 4000. On this picture membranes were exposed up to 20 mins. Transfer by the way was great. I can email you the picture with the tranfer. I just took a picture of transferred membrane just in case. I hope you can help me with this. Thank You
Asked on Dec 02 2011
Thank you for keeping me updated about this situation and for sending the protocol information and image. I unfortunately can not open the Gel file. Could you re-send it in pdf or jpg? I will be happy to see if there are any suggestions that might improve the results. The replacement antibody is also covered by our Abpromise so we would be happy to issue a credit or refund if we can not improve the results. Please let me know if you have any questions or if there is anything else you need.
Answered on Dec 02 2011