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Customer is seeing only a diffuse band at the expected size of 15 kDa on rat ganglia samples and a strong band ~30 kDa. Loading 20ug sample, running tubes for 10 minutes on denaturing program on PCR machine. Using 1 ug/ml for staining with primary. Question for lab: How was tissue solubilized?
Asked on Oct 31 2011
Thank you for calling today, it was a pleasure speaking with you! As we discussed over the phone, your protocol sounds reasonable and you are using the antibody on tissue we have tested in-house. I'm unsure of why the antibody is not working on your samples, and apologize for the difficulty you've encountered. As we discussed, checking the denaturing program might be a good idea just to ensure that you're fuly denaturing the proteins in the sample to prevent any dimers or complexes. I have arranged for you to receive a free vial of a different lot of the antibody on order number ######. This should arrive to you tomorrow. Please let me know how you fare with the new vial, I'll be curious to know if the new lot makes a difference! If you have any other questions, please do not hesitate to contact us. Have a great evening!
Answered on Oct 31 2011