Here are some questions I have when a customer has a problem with a western blot with no bands and over-expressed lysate with RT-PCR confirmation.
-What was the protein concentration of the cell lysate when loading on the gel?
-Can you confirm the over-expression method? Was it the full-length sequence?
-Have any proteomic methods confirmed the over-expression of the target? Is it possible this is a limit of detection issue?-Did another antibody work for this application?
-Would your customer be interested to test another antibody of Aviva?These answers will help us determine if there is a Quality Control issue with this product.
Thanks Matt