Anti-Histone H2A.Z (acetyl K4 + K7 + K11) antibody - ChIP Grade (ab18262)

Dear tech team,  A customer I spoke with recently would like to have a bit more information on the cross-reactivity of some antibodies they have bought: ab 18262: H2A.Z (acetyl K4+K7+K11) ab18263: H2A.Z ab4174: H2A.Z ab18255: H2A  Could you please let me know if you have performed any dot blots or ELISA with different recombinant histones or recombinant histones with post-translational modifications of the histones.  It is very important for them to know that the Abs don't crossreact with other histones (H2A, H3, H4, H1, H5) and histone modifications, very important for us is to know if they pick any acetylation (H3K9 H3, H2Ak5, etc) or methylation (H3K27, H3K9 ETC).  Could you please let me know if you have this information available?  

ANSWER:

Thank you for contacting us.

Unfortunately, these H2A.Z- and H2A antibodies have not been tested in ELISA or dot blots and we therefore cannot guarantee them to work in these applications.

Ab18262 and ab18263 have been tested in Western blot against recombinant H2A.Z as well as with endogenous histones, whereas ab18255 has also been tested against recombinant histones H2A, H3 and H4 and endogenous histones, as has ab4174 against endogenous histones (e.g. calf thymus histone lysate).

All these testings as well as our customers' feedback in Abreviews demonstrate that in Western blot these antibodies are specific for H2A and H2A.Z, respectively, and do not recognise any other histone. For ab4174 cross-reaction to histones H1 and H5 was observed, the reason for which is unknown. However, this only occurred in AUT gels and never in standard Western blot.

Furthermore, ab18263 was shown to not recognise acetylated H2A.Z nor any other acetylated histone, whereas ab18262 detects only the acetylated H2A.Z and does not recognise the non-acetylated form. ab4174 and ab18255 may detect modified histones, however, they have not been tested against any histone modification.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

I am very interested in your product, H2A.Z (acetyl K4 + K7 + K11) antibody - ChIP Grade (ab18262). I wonder if your antibody recognize H2AZ that is acetylated on K7 and K11, but unmodified on K4.

ANSWER:

Thank you for your enquiry.

It's not possible to be absolute in answering this question. The immunogen was an N-terminal peptide acetylated at all 3 lysines and affinity purified against the immunogen. When Westerns were used to screen the serum against total HeLa butyrate histones and immuno-precipitated DNA, three bands could be seen - which presumably represent mono-, di- and tri-acetylated H2A.Z. This means that partially acetylated species are recognized. However, the precise nature of these partially acetylated species remains unknown, i.e. the site occupancy is undefined, so to what extent H2A.Z acetylated on K7 and K11 but unmodified on K4 is a component of the di-acetylated state is unknown. The serum has not been tested against a range of partially acetylated N-terminal peptides - which would probably be the best approach to answering this question.　My guess is that H2AZ acetylated only on K7 and K11 would be recognized.

I hope this information will be helpful. If there is anything else that I can help you with, please do not hesitate to contact me.

Bonjour,

We've been notify by Lyne St-Pierre (our purchasing responsible) that the order reference number 138584 for the antibody ab18262 was cancelled due to USDA/Canadian restrictions. Can you give us more details cause it is only an antibody!? Do you have another antibody that recognized an acetylated form of mammalian H2A-Z (Chip grade preferentially)?

Thanks,

ANSWER:

Thank you for getting in touch with me. I am sorry for the confusion. Produce derived from farmyard animals cannot be lawfully shipped directly into the US or Canada. Unfortunately this includes chicken and sheep antibodies.

Unfortunately we do not have an alternative acetylated histone H2AZ antibody available in our catalogue.

Please let me apologise for this once again.

I am interested in your products, ab18262 and ab18262 (antibodies for histone H2A.Z), but I was told that these idems are US restricted. Could you plaese tell me the reason and how I can get those antibodies?

ANSWER:

Thank you for your enquiry.

US Customs are currently preventing all antibodies raised in sheep, goat, donkey, horse, pig and chicken from entering into the States unless you have a USDA import permit in place for these items.

You can view information on import restrictions at the UDSA Animal and Plant Inspection Service information page: http://www.aphis.usda.gov/vs/ncie/biofacts.html

Abcam’s US office in Cambridge MA holds an import permit that covers for the majority of antibodies in our range that are derived from farmyard animals outside the USA. Unfortunately our permit does not cover this product and thus we are unable to ship it to our customers in USA.

I hope this information helps, please do not hesitate to contact us if you need any more advice or information.