Anti-Histone H3 (acetyl K18) antibody - ChIP Grade (ab1191)

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ANSWER:

Beide Artikel wurden in Großbritannien hergestellt und haben ein Gewicht von 100ug.

last july I ordered some antibodies. I can't find anywhere what I should use for a positive control. Please can you tell me

We use this antibodies for parafine section of breast cancers

ab1191 - h3 (acetyl K18)

ab1761 - h4 (acetyl K12)

ab4441 - h3 (acetyl K9)

ab7766 - h3 (dimethyl K4)

ANSWER:

Thank you for your enquiry.

Details of the positive controls for these histone antibodies are not detailed in the datasheet. However, these histone modifications are ubiquitous across all cell types (except yeast which do not have acetylation at lysine 12 of H4). You could use a standard HeLa cell line and prepare them as detailed in the abcam immunocytochemistry protocol (http://ops.abcam.com/index.html?pageconfig=view_protocol&pid=159)

or alternatively,

http://ops.abcam.com/index.html?pageconfig=protocols&pid=174&intAbID=7766&strTab=protocols&mode=prot

It is worth noting that you can elevate the level of histone acetylation across all of the lysine residues you have antibodies against by incubating (if possible) with Na Butyrate (5mM). This will in turn improve your levels of staining.

Please do not hesitate to contact me should you require further assistance.

Thankyou for your reply. I am aware of the effort Abcam is putting in testing their antibodies. I am very happy to learn that Abcam is continuing to improve its quality controls and hope my data were of constructive help in this process. As for the refund: The antibody ab1191 had been ordered the 12th of may 2005 from ICMB stores for my group (Vogelauer) with the order number: MB54303MV. I hope I will have more positive reviews for your website in the future,

ANSWER:

Thank you for your order details.

I am sorry that we were not able to improve your results to your satisfaction. I have asked our accounting department to issue you a credit note against purchase order number MB54303MV (Abcam's purchase order number 83242). I hope that this experience will not prevent you from purchasing a product from us in the future as we have many quality products.

The technical team is always at your service, should you require further expert advice.

Good luck with your research.

thankyou for inserting my corrections to the abreview. As a general remark - I would have appreciated to see your writing BEFORE putting it on the web together with my data. I appreciate the point-system of Abcam as encouragement to comment on their products at your web-site, although I think that in this case a refund would be more appropriate, as I received a product that didn't correlate to what it was advertised for. (Let alone the effort to come to this conclusion.) I was also curious if Abcam is planning to change the way of testing their histone modification antibodies. Currently Abcam is testing their antibodies for the ability to Chromatin-immunoprecipitate DNA, but unfortunately in a way that does not reveal anything about specificity. Why not taking advantage of yeast genetics? As histone-tails are highly conserved, these data would still be valid for other organisms. The relevant yeast strains exist and I am sure that research-labs would be happy to provide them to companies for testing-purposes. Positive and negative controls have already been established. So the working effort of Abcam would not change, but the information they could provide would be so much more valuable for their clients. And I would think that - given the choice - any researcher would option for the antibody who's specificity he can rely on, without the need of time consuming tests. Finally and most importantly, a not accurately tested product can lead to a lot of confusion in the literature, as data are pubblished under mistaken premisses. I hope Abcam will consider to update its way of testing the specificity of histone modfication antibodies in ChIP experiments. I am sure that the entire chromatin field would appreciate such an effort very much.

ANSWER:

Thank you for your reply. We submitted your review on your behalf because we did not want to miss our on such great feedback with such carefully controlled experimental data.

The policy at abcam is to test the specificity of our antibodies using ELISA or western blot. I appreciate that this does not verify the specificity under ChIP conditions. I also acknowledge the advantages of employing yeast with point mutations at the modifiable lysine/arginine residues. Our current policy is to first examine specificity using ELISA and western blotting, then test whether the antibody can precipitate under fixed conditions using ChIP. Only then is it referred to as ChIP grade. However, I do appreciate the importance of the site specificity of our antibodies and how researchers have come to rely on our datasheets. I also understand the implications to the chromatin field given the importance of the pattern of histone modifications and the epigenetic code.

We do have measures in place to perform a more detailed examination of the specificity of this antibody in future batches, albeit not currently by ChIP.

Please can you e-mail me your original purchase order number so that I can arrange for a refund.

Charlotte Your antibody ab1191 works well on mouse tissues, prostate, uterus and lung, following citrate retrieval, tho' at the moment it does not look as if we will be pursuing this line of research.

ANSWER:

Thank you very much for getting back in touch with me and for the feedback that you have offered. I would encourage you to submit a review relating to the application that you tested the Ab in and be rewarded for your feedback.

In return for a review we will offer you 50 “Abcam Points” per a product and separate application reviewed. Abcam points may be redeemed on a number of rewards (a further 100 points will be offered for an image).Ab1191 may be reviewed via the link at the base of the on line data sheet. If you have any questions relating to this process then please get back in touch with me.