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

Our customer purchased 1 vial of ab6741 and 1 vial of ab6721 last December when we had some promotions.
He stored it 4C and openend it this morning to test.
But there were severe precipitation in the vial (ab6721, Lot.GR27131-6, ON.1005934).
I recommended to use after spin down but,it alreadyoccupied much portion of the vial,
he doesn't want to use it because hethinks he lost the volume about what he purchased 500ul of ab6721.
Please arrange a replacement vial for our customer.
Best regards,

ANSWER:

Thank you for your email. I am sorry to hear that you are experiencing problems with these antibodies.

The brown precipitates with HRP conjugated antibodies are very common. We recommend customers to pipette up and down and vortexing the vials which actually helps in dissolving the precipitates. As a final measure we also recommend centrifuging out the precipitates. It is true that it will affect the volume however because the product is at high conc. the effect will not be huge, only few mictolitres will be lost.

Please do not hesitate to use the antibodies in applications as recommended on the datasheet.

Please also note that we recommend storing this product at 4C for 1-2 weeks only, then it should be stored at -20C. The customer have stored the antibody at 4C since December 2011 which actually voids the Abpromise guarantee. Unfortunately replacement vials or refunds cannot be offered. However we are happy to troubleshoot he problem in case these products does not work.

I hope this information will be helpful. Should you have any question please do not hesitate to contact me.

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.