Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220)

I've been corresponding with Abcam technical support about ICC trouble with ab1220. I still can't get it working. I would like a replacement with ab8284.

ANSWER:

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with ab8284 with the order number 1088925.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

Hello,
I am a little confused about my antibody lot number. I think a different lot number is printed on my tube compared to the lot number on the paperwork that arrived with the antibody. Is lot# 1169286 the same as lot# GR59571-1 for AB1220?
Best Regards,

ANSWER:

Thank you for your enquiry regarding ab1220.
I can confirm that GR59571-1 is identical to batch: 1169286. These two numbers refers to the same purification.
I am very sorry for the confusion caused!
I hope this helps and if I can assist further, please do not hesitate to contact me.

Great thanks for this. But just to check. On the datasheet for the histone peptide that you reference ab1772, it suggests its partner antibody to the peptide is mAbcam62168, a seemingly different antibody to ab1220, the one I've requested. Just want to be certain that they share the same immunogen and if so, maybe you could explain what makes these two antibodies different then.
Many Thanks,

ANSWER:

Thank you for getting back to me.
You are correct in thinking the peptide ab1772 can be used as the blocking peptide for both antibody ab1220 that you have been using, as well as ab62168. The same peptide was used to raise both of these monoclonal antibodies.
The antibodies are different only in that one is HRP conjugated (ab62168) and the other is not (ab1220). I realise that this may be a little confusing as we have given the different antibodies different clone names and I will see if this information can be updated.
If you have any further questions, please do not hesitate to contact us again.

We had corresponded previously about the Ab5103 antibody and I
want to thank you again for resolving that for us. I have a query about
another histone antibody, Ab1220. We have used it a couple of times now
and are getting very weak staining, which is showing up in both the
cytoplasm and the nucleus. We were wondering if you could recommend a good
positive control for this antibody (preferentially mouse cells) and if
anybody else has reported such a problem.
Best Regards,

ANSWER:

Thank you for your email; it's good to hear from you again. I hope you have been doing well and your research has been progressing. Did the most recent vial of ab5103 produce the expected results?

Regarding ab1220, I am sorry to hear about the problems with this antibody. I've looked through our records, and we haven't receivedany otherreports of weak cytoplasmic and nuclear staining. For ICC, we test the antibody on HeLa cells, however other researchers have reported good results in a mouse hepatoma cell line and SV40-transformed MEFs:

http://www.abcam.com/index.html?datasheet=1220&tab=abreviews&intabreviewid=9406

http://www.abcam.com/index.html?datasheet=1220&tab=abreviews&intabreviewid=8089

I'm not sure that either of these murinecell types will be easy to obtain, so I might recommend using the HeLa cells, at leastinitially. Have you tried the antibody in any other applications, like Western blot? What kind of mouse cells are you staining?

If you would like to forward any protocl information and data, I may be able to suggest some alterations to improve the results. I'd also be happy to send a different lot if you'd prefer.

I look forward to hearing from you. Please let me know if you have any otherquestions or if there is anything else that we can do for you, and I'll be happy to help.

Product code: 1220
Lot number: ******

Inquiry: I am trying to figure out the specificity of the H3K9me2 mAb (ab1220). Will you be able to send the data for your blocking/competition experiments?

ANSWER:

Thank you for your patience while I contacted the lab to get the specificity data that you were looking for. Please find all of the data we have on ab1220 in the attached pdf.
Please let me know if there is anything else I can help you with.