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Please see our reponse. Thanks.
1 - What sample type are you using? : we tried 293T cell lysates and mouse tissue (liver) lysates prepared in RIPA buffer.
2 - Are you seeing any bands, no bands etc?: no clear bands, there are a few very faint ones after long exposure
3 - How much sample are you loading? : we tried from 20 to 100ug per lane
4 - What type of blocking agent are you using? : we use 5% non fat dry milk in TBST.
5 - What primary antibody concentrations have you tried and how long do you incubate for?: we tried 1:500 ˜ 1:1000 in 1% BSA in TBS, incubated overnight (16hr) at 4 degree with rocking motion.
Asked on Jul 26 2012
Thank you for your reply.
I am sorry that the antibody is not working for you an based on the information that you have provided, I do not believe that there is ay protocol advice that I can offer that would resolve the issue.
Therefore I would like to offer either a replacement antibody or a refund. A possible aletrantive could be:
Please let me know how you would like to continue.
I look forward to your reply.
Answered on Jul 26 2012