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Here is the lysis buffer ingredients list:
Asked on Aug 28 2012
Thank you for taking the time to provide these details. Theinformation you have kindly provided will enable us to investigate this case for you and this is also helpful in our records for monitoring of quality.
Reviewing this case, I would like to offer some suggestions to help optimise the results. I would also appreciate if you can confirm some further details:
1. There are inherent difficulties with antibody detection of recombinant proteins that I can recommend considering, particularly if they are tagged. This may be a biological artifact rather than a problem with the antibody.Could you confirm that the protein transfected is full length and includes the immunogen sequence? Also, if the recombinant TC10 is tagged, it is possible that the tag is preventing access to the antibody. Could you confirm at which region of the protein the tag has been placed?
2. Could you confirm the species from which the recombinant TC10 gene was taken?
3. Could you confirm how the stability of transfection has been assessed?
4. I can recommend that due to the points described above, it is important to use an endogenous positive control sample. For example, this antibody has been used successfully on293 cell lysate.
I hope this information is helpful, thank you for your cooperation. Should the suggestions not improve the results with an endogenous positive control, please do not hesitate to contact me again with the further requested details.
Answered on Aug 28 2012