Anti-pan methyl Lysine 抗体 - ChIP Grade (ab7315)

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 the order number XXXXX. 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.

As both antibodies failed to work in western blot, I have sent you ab23366 as a free of charge replacement for ab7315, your new order number is ******** andI have also credited the cost of ab412, your credit note number is **********.

I am sorry about the problems you had with these antibodies and I hope that ab23366 works out for you. If there is anything else I can do, please let me know.

Hope you have a good evening.

I really appreciate you sending that further data.

Since it is clear that ab412 and ab7315 are not detecting your protein of interest in your western blot and under our Abpromise, we guarantee that both should work, then I would be very happy to either send you replacement antibodies or to process a refund.

Please let me know how you would like to proceed and if you would like to receive replacements antibodies, let me know the catalogue numbers of those you would like. Could you also provided me with the original order details for these antibodies (either order #, purchasing agent name, date of purchase) so that I can complete your request.

Thank you once again for your patience and I look forward to your reply.

Thank you for answering those questions and your continued patience.

Firstly, I would just like to reiterate that both ab412 and ab7315 are guaranteed to work in IP and so if we cannot determine what the problem is, then I will be happy to replace or refund these antibodies.

Having said that, I do have a few more questions/suggestions:

1 - Have you confirmed that the Myc antibodies are actually IP'ing your sample of interest. You could determine this, buy using the Myc N262 to IP and then blot with MycC33.

2 - Your input sample (you load 20ug?), does give a signal for your protein of interest, but have you tried starting with more total protein. I ask this as although you are using 20ug of total protein but increasing that amount (to say 40ug) you may be able to get more of your protein of interest to actually be IP'd and so get a signal using the antibodies.

I look forward to your reply and helping you resolve this issue.

Thank you for providing that extra information.

Just to make sure that I understand your western blots:

1 - You are using a Myc antibody to do the IP, correct?

2 - What is the size of your band of interest? As you are seeing a band about ˜60kDa in your inout lanes and also seeing the same size band in your control and IP lane.

3 - Are you only expecting to see bands in your IP lane, NOT your control IgG lanes?

Thank you for your patience in this, I just want to make sure that I understand the problems that you are having, so that I can give you the best advice or send you the appropriate antibody.

I look forward to your reply.

Thank you for contacting Abcam.

I am sorry that you had disappointing results using some of our products, under our Abpromise, we guarantee our products to work as stated on the datasheet. Since you have been having trouble with them, I will be happy to look over protocol that you have been using and see if there is any suggestions I can make. Could you please tell me the types of samples you are using, application antibody is being used in and also the general protocol you have used (including primary antibody concentration, incubation times, blocking buffers etc). Also, could you provide a description of the problem that you are having, or if you can send a copy of the results that you are seeing. We do take all complaints about our products seriously and do our best to resolve these issues.

I look forward to your reply and helping you resolve these issues.

Thank you for your message. You mentions ab80182 as well as ab76118 at the beginning of this email. I assume you are meant to refer to datasheet of ab76118 only, not ab80182? I apologise for any confusion. I have been in contact wtih our source in order to clarify this information. They have confirmed we were regrettably originally sent incorrect information for the previous enquiry. I can confirm that this antibody will not cross react with acetylated proteins, mono-methylated proteins, or di-methylated proteins. We aim to provide accurate and correct information to customers, and I am sincerely sorry there was some confusion in this case. I have removed the published enquiry that had the incorrect details on the datasheet. With regards to ab7315, I am sorry this did not come up in my previous search on our system for you. I can confirm that this is tested in IP, and wil be suitable for this application. Thank you for your understanding. If you have any further questions, please do not hesitate to contact us. 

Thank you for your reply and for confirming the product numbers. I am pleased to provide the following information: ab76118 Methylated Lysine antibody: This antibody specifically recognizes proteins that contain trimethylated lysines. It will cross react with acetylated proteins, mono-methylated proteins, or di-methylated proteins ab7315 pan methyl Lysine (methyl K pan) antibody: This antibody has been tested with tri-methylated lysines (K4, K27 and K9) but it was found that the antibody recognizes these to a lesser extent than di-methylated lysine residues. This information is available on the datasheet underneath the western blot image. There is also a note in the specificity section stating that histone H3 tri-methyl K27 was recognized to a lesser extent in ELISA analysis. I hope this information will be helpful to you. Should you have any further questions, please do not hesitate to contact us.

Thank you for contacting us. I am very sorry to hear you are still having a problem with ab7315 (pan methyl Lysine (methyl K pan) antibody - ChIP Grade). I'm also sorry I didn't ask earlier but could you clarify a couple of things for me: 1) You mentioned: "Bacterially expressed and purified GST-tagged protein (of human origin) which has been methylated in vitro using the methyltransferase SET 7/9" as well as "However, the antibody picked up protein whether it was methylated or not, suggesting that this low level binding was non-specific" but doesn't methylation take place in bacteria - are you using a mutant strain? Have you tried in vitro translation of histone - can you do that? 2) You mentioned: "Incubation overnight with the antibody improved the signal (but only on an overnight exposure to film). However, the antibody picked up protein whether it was methylated or not, suggesting that this low level binding was non-specific. did you detect improved signal with increased protein loading or no signal at all? I have confirmed your Abcam order 183065 (1X ab7315, lot 166243). Once I receive your reply and determine there is a problem with the antibody, I will be happy to replace ab7315 with ab23366 for you. I apologize for any inconveniences and I look forward to receiving your reply.