Anti-N epsilon gamma glutamyl Lysine 抗体 [81D4] (ab424)

Thank you very much for keeping me updated.

I'm glad to hear that the results seem to have improved. We currently have only that lot in stock (2 vials in our US stock and 1 vial in our UK stock), so there should be consistency in results.

Please let me know if you have further questions or need anything else, and I'll be happy to help.

Thank you very much for your reply.

I'm sending a free of charge vial of ab97230 on the order ***, which should ship out tonight and arrive tomorrow.

I've also included the promo code for the cell imaging book.

Please keep me updated about the results with this secondary antibody, and let me know if you need anything else.

Thank you very much for your reply.

For your assay, either of these antibodies will be suitable. I would suggest ab97230 because it is in stock and we can ship it to you overnight. Ab98679 is back-ordered and I expect it to work similarly to ab97230.

I'd be happy to send this to you free of charge if you would like. Please let me know your order information and I will set this up for you.

Thank you very much for your reply.

I apologize but we won't be able to test the antibody with your samples in our lab at this time. I'm very sorry for the inconvenience. Did you check whether the mouse-on-mouse HRP polymer detection kit works with mouse IgM isotype primary antibodies? If you'd like to send me the catalog number of the kit you're using, I'll be happy to look into that for you.

We only have the same lot in stock that you're currently using, but we can still send a replacement vial to see if the results improve. If you'd like to try this, please send me your original order or PO number and I'll arrange this for you.

I am sorry that I don't have more to offer at this time, but please let me know if you have any questions or need anything else and I'll be happy to help.

Thank you very much for keeping me updated about the results.

I'm sorry to hear that altering the protocol did not help improve the results. Have you also tried staining sections without doing antigen retrieval at all? I apologize that we don't have an in-house IHC-P protocol for this antibody, as it has only been tested in this kind of tissue staining according to the literature. Are you certain that your secondary antibody or detection system is reactive with mouse IgM isotype? An anti-IgG secondary will probably react with the conserved light chains of the IgM molecules, but this wouldn't be as strong as an anti-IgM staining.

What lot number are you using right now? We have lot a new lot in stock, and I'd be happy to send you a free of charge vial to see if the results are different. If you'd like to try this, please let me know your original order or PO number and I'll be happy to send this to you.

I look forward to hearing from you. Please let me know if you have any questions or need anything else and I'll be happy to help.

Thank youvery much for your reply and for sending these additional details.

I have a couple of suggestions that might help improve the results. It may be useful to decrease the antigen retrieval time to 15-20 minutes, in order to reduce the reversal of cross-linkages. Do you know if the rodent decloaking agent is similar to citrate buffer at pH 6, which has previously worked in antigen retrieval with this antibody? I'd also recommend incubating the antibody overnight at 4C instead of 2 hours at room temperature.

If these alterationsare not helpful in improving the results with this antibody, I'dbe happy to send a free of charge replacement antibody orissue a credit or refund. Please keep me updated about any further results, and let me know if you have any questions or there is anything else that we can do for you.

Thank you very much for your reply, and for your patience.

Thank you also for sending the images of your staining results. I've reviewed some of the literature in which this antibody was used in IHC-P, and there aren't many more details about the protocols used. Have you tried increasing the incubation time, such as overnight at 4C if you're currently incubating for 1 hour at room temperature? What kind of blocking solution is being used? I'd recommend blocking with 10% serum for up to an hour at room temperature. What kind of secondary antibody and detection system is being used? Could an amplification be done (avidin-biotin complex, etc)?

If none of these ideas are useful, then I'd be happy to send a replacement vial or issue a credit or refund. Please keep me updated with any new results, and let me know if you have any questions or if there is anything else that we can do for you.

Thank you very much for your call last week and for your patience while I've looked into your enquiry.

We have not tested this antibody in our own labs in IHC on formalin-fixed paraffin-embedded tissue, but it has been testedin IHC-P according tothe literature. One publication by Watts et al indicated that they used an Unmasking Reagent from Vector-

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2682564/

In another article by Tovar-Vidales et al, 10 mM citrate buffer pH 6 was used for antigen retrieval-

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2648869/

I haven't found any references showing that antigen-retrieval was not used, so we don't know if the antibody works better without this step. If you'd like to send some more details about your protocol, I'll be happy to review it and see if there are any suggestions to improve the results. We do fully guarantee this antibody to work in IHC-P, so we can send a replacement or offer a credit or refund if the results are still not satisfactory.

I look forward to hearing from you. If you have further questions or need anything else, please let me know and I'll be happy to help.

I can confirm that this antibody will detect N epsilon gamma glutamyl Lysine also if part of a human protein. In the publication PubMed ID 18235007 for example, researchers have used this antibody successfully in human paraffin embedded tissue slides. We actually have not stated species reactivity on the datasheet, as N epsilon gamma glutamyl Lysine is a very small epitope and its recognition should therefore not be dependent on the species.