Question (45934) | Anti-SCAP antibody (ab125186)

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Question

Hi,
I am writing in regard to your anti-Insig2 antibody (ab86351), which hasn't been working for me at all. Attached is the western blot of one of my experiments, where I expressed FLAG-HA-tagged human Insig2 in HEK 293T cells, and blotted with anti-Insig2, anti-FLAG or anti-HA antibody. As you can see, anti-FLAG and anti-HA antibodies detected the tagged-INSIG2 pretty well, while no specific band showed up in the anti-Insig2 blot. I have also tried this antibody on rat and mouse liver lysates as well as HepG2 whole cell lysates and it could not detect the endogenous Insig2, either. I was wondering whether you could send me your other anti-Insig2 antibody (ab86415) as a replacement.
Besides, I was wondering whether you could send me some samples of your anti-SCAP antibodies (ab125186 and ab91323). I would be happy to post my blots if the antibody works.
Thanks,

Answer

Thank you for contacting us and for letting us know about the trouble with ab86351. I am sorry to see that the results have been so poor.

I would be happy to send a vial of ab86415 as a replacement. Do you have your original order or PO number, so that I can arrange for this replacement?

For our records, I also have some additional questions about the protocol:

1) How much protein was loaded per lane of the gel?
2) What protease inhibitors were used, and was this a standard reduced and denatured Western blot?
3) What was used to block the membrane (i.e. milk or BSA)?
4) What dilution of antibody was used?

We only have full size vials of all of our products, so unfortunately I am not able to send samples ofthe SCAP antibodies ab125186 and ab91323. We do guarantee them both to work in Western blot with human samples, so if you had any trouble with them we would issue replacements, credits, or refunds. If you would be using them in an untested species (like mouse or rat) we do have a testing discount program, and I would be happy to send more information about that program if it's applicable.

I look forward to hearing from you and resolving this promptly. Please let me know if you have any questions or if there is anything else that we can do for you.