Question (28817) | Anti-V5 tag antibody (HRP) (ab1325)

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Question

Hello, I am unsure if this is the correct e-mail address to send my concerns, if it isn't could you please forward this to the correct department. I recently ordered Anti-V5 tag antibody (HRP) product number ab1325. I have tried numerous different times to use this anti-body to detect a recombinant V5 tagged protein. On each attempt I have ended up with either a very high level of non specific membrane staining which makes it impossible to determine the level of recombinant protein, or a large amount of non specific bands which also make it impossible to determine the level of recombinant protein. I've tried over night blocking in both 5% fat free milk or 5% BSA and also 1hour blocking in % fat free milk. Between 5-10, 10minute washes in 0.1% PBS-Tween. And a series of dilutions with 1:5000 used the most. I have compared your antibody to a HRP-V5 tag antibody from Invitrogen, which gives a single band at the predicted mass on the same protein samples. Unless there are some very specific conditions which are needed to use this anti-body I am unable to get it to perform in a tested application as described on the datasheet. Would you be able to give me details on either a refund, or a replacement product. Regards,

Answer

Thank you for your enquiry. I am sorry to hear this vial of ab1325 is giving you trouble. It does appear that this antibody is not detecting the V5 tag as expected. If you have purchased it within the last 6 months or so I would be happy to offer you a free replacement, credit or refund. In your reply, please include the purchase order number or Abcam order reference number associated with the initial order of ab1325 so I can process your request. Thanks! I look forward to hearing from you.

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