Anti-EHMT2/G9A 抗体 (ab40542)

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Thank you for contacting Abcam.

I am sorry about that you are experiencing some problems with ab40542 in ChiP. The antibody is covered under our Abpromise for six months and is guaranteed to work in ChIP on human samples . If we cannot resolve the issue you are having with the antibody then I would be happy to either send a replacement antibody or to process a refund.

When you say that the antibody did not work as well as previous lots, were you still getting a signal using the current lot of ab40542, but just had to use a higher concentration of the antibody, or did the antibody not produce the expected results at all. Also, did both of the antibody vials you received have the same issue?

Would you also be able to provide the order or PO number used to purchase these vials of ab40542, so that I can locate your order details.

I look forward to your reply.

Thank you for contacting us.
For ab40542: As immunogen, the recombinant fusion protein, corresponding to amino acids 831-1001 of Human G9a was used to raise the antibody. The epitope has not been mapped.
As for the DNMT1 (DNA methyl transferase 1) antibody, coud you please let me know which antibody (catalog number) you are refering to? Thank you.
I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.
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Thank you for your reply. I am sorry that this antibody is producing poor results in WB. Based on the images provided, I suspect the antibody has lost specificity with age. It is possible that the results may be improved with troubleshooting, but it would bechallenging given the intense background andweak specific band. The following may help:

1)Given that KMT1C is a nuclear protein, I would recommend preparing the lysates with RIPA buffer. The specific band may be strengthened and the background reduced by enriching the samples in a nuclear extract. Sample degradation could account for some of the background bands, so fresh lysates prepared on ice with protease inhibitors would be recommended.

2) What blocking agent was used? This antibody was validated using an overnight block with milk, which is recommended for keeping the background low.

3) How much primary antibody was used and how long was it incubated for? We recommend using a 1:2500 dilutionfor 2 hours at room temperature. If you continue to seebackground at thisdilution, you could try reducing the secondary antibody concentration.

I hope this helps, please let me know if you need any additional information or assistance.