Rabbit polyclonal Secondary Antibody to Goat IgG - H&L (HRP) (ab6741)

Thanks for your reply. I did get some non-specific binding with the secondary only control but not as high as with both primary and secondary. I am using a very sensitive ECL reagent that we are developing and I believe I get the background in the secondary only control because I am doing long exposures to try and see the specific signal.

I would appreciate a replacement for the Primary antibody and I thank you for your help in resolving the matter.

ANSWER:

Thank you for your reply.

I was wondering if ab8227 would be a good replacement for you:

http://www.abcam.com/beta-Actin-antibody-Loading-Control-ab8227.html



If it is, then please let me know and I will get it sent out to you as soon as possible.

I recieved ab7125 as a replacement for ab6741 and tried it. I still have the same problem of non-specific background binding. I still believe as I originally thought that the primary antibody ab8229 may not be working. I have run controls where I use purified actin (from Sigma) and even though I can see some bands, the signal to noise ratio and detection limit indicate that this is from non-specific binding of the secondary antibody. I can send you a scan of the film if you would like.

ANSWER:

Thank you for your reply.

I am sorry the replacement secondary did not solve this problem. Do you get the same non-specific binding pattern when you do your secondary only control? I ask, because if you do, then that does indicate that the issue is with the secondary, not the primary (as was the case with the first secondary antibody you received).

If however, you are not seeing the non-specific binding in your secondary only controls, then I agree that the primary antibody is the problem and therefore I would be happy to send another one free of charge to resolve the matter.

I look forward to your replay and helping to resolve this issue.

I would be happy to try the replacement.

Thanks.

ANSWER:

Thank you for your reply.

As ab6741 did not prove to be successful for you, I am sending you ab7125 as a free of charge replacement. Your ne order number is ******* and you should have the new product by 10.30am tomorrow morning.

If there is anything else I can help you with, please let me know.

Regarding product ab6741, I did change the blocking reagent to BSA with no success. There was still high background. I also tried using blocking agent at every step, i.e. I put the blocking agent in with the secondary antibody. In that case I got no signal.

The antibody ab6741 was ordered on*******with PO number *******.

ANSWER:

Thank you for your reply.

I am sorry that you are continuing to have problems with ab6741 in western blot. Therefore I would like to send you ab7125 as a free of charge replacement:

http://www.abcam.com/Donkey-polyclonal-Secondary-Antibody-to-Goat-IgG-HL-HRP-pre-adsorbed-ab7125.html

Please let me know if this is ok with you, or if you would prefer a refund and I will be happy to have it sent out as soon as possible.

Looking forward to your reply.

High background from the secondary antibody.

ANSWER:

Thank you for contacting Abcam earlier today.

If you are still experiencing any issues with ab6741 in western blot after trying the protocol tips that we discussed, please let me know and I will be happy to look into sending a new antibody to try out.

Please let me know if there is anything else I can help you with.