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Thank you very much for your help.
I'm afraid the person who ordered this antibody has since left and we do not have access to the order details. However, we do know that it arrived in the lab on the 4/10/11.
I have attached the questionnairedetailing the western blot protocol used.
Hope to hear from you soon.
Asked on May 22 2012
Thank you for sending the questionnaire.
It is difficult to guess, the identity of band at 30kDa which is consistent in all lysates. The only lysate which shows correct band size is LMX1B trans-fected Hek 293 cells. In regards to other lysates I am sorry we are unable to determine the identity of extra bands; you may need further optimization of the application. My suggestions for optimization would be
- Include negative control i.e. lysates without primary antibody
- Prepare lystes with 80-90% cell confluency
- Keep the lysis buffer at 4C when lysing the cells
- Boil the lysates in sample buffer for 10 minutes at 100C.
Please note that the purchase of this antibody is out of Abpromise guarantee. However we are happy to help as far as we can.
I hope these suggestions will help to improve results. If these doesn't please let me know, I will then try to replace the antibody free of charge.
Answered on May 22 2012