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Product code: 39155
Lot number: gr3280-1
Inquiry: I've been using this antibody to validate a mass spec experiment whereby ADAM12 was identified as a potential target. We have performed RT-PCR to confirm its expression. I brought this antibody to use for validation primarily by flow cytometry as the data sheet supports the use of this antibody by flow and it also recognises an N-terminal extracellular epitope within the protein. However i have failed to get this antibody to work by flow cytometry despite numerous attempts on multiple human cell lines (all of which are positive by mass spec and RT-PCR). I have tried using trypsin and cell disociation buffer (preserve cell surface epitopes) for cell release from flasks and have used either no blocking (PBS alone) or FCS or BSA as a blocking agent. I have always run the flow using other positive markers that require the same rabbit secondary antibody with positive results. I have tried using down to a 1:50 dilution of the antibody (suggested is 1:500) with no change in result. As the data sheet suggests it can also be used by western blotting i have also tried using the antibody for this purpose twice using a concentration of 1:1000 or 1:500 and 5% non-fat milk for blocking. Again i have achieved no positive staining, although probing for other targets alongside ADAM12 with an identical rabbit-HRP secondary have been successful. Reading the data sheet i was a bit concerned by the size of the product quoted for the shown western blot as the predicted band size is 100kDa, but the shown blot has a major size band of 42 kDa and minor bands at 50 and 84kda. Please could you advise what the next steps are or if you have an alternative antibody that might be more appropiate?
Asked on Jul 12 2012
Thank you for taking time to complete our questionnaire and for contacting us. I am sorry to hear this antibody is not providing satisfactory results.
The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality. I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful.
Having reviewed the protocol details, I believe this product should have given satisfactory results. It appears that you may have received a faulty vial.
I apologize for the inconvenience and am pleased to offer you a free of charge replacement, credit note, or refund in compensation.
Thank you for your cooperation. I look forward to hearing from you with details of how you would like to proceed.
Answered on Jul 12 2012