Anti-Histone H2A.Z antibody - ChIP Grade (ab18263)

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.

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DESCRIPTION OF THE PROBLEM Hello, Can you please tell me if any of your H2AZ antibodies (ab18263 and ab4174) are predicted to react with Drosophila melanogaster?

X-CHIP or N-CHIP Hello, I would like to know if the H3 antibody recognizes both H3 and H3.3 or if it is selective just to H3 (in Drosophila) ?

ANSWER:

Thank you for your enquiry.

I have performed an alignment with the sequences of each of the immunogens of ab4174 and ab18263 against Drosophila sequence NP_524519. Once again the results can be viewed below.

http://www.ebi.ac.uk/cgi-bin/clustalw/result?tool=clustalw&jobid=clustalw-20051219-10413321&poll=yes

There is perfect conservation of the immunogen sequence of ab18263 in the Drosophila sequence. I have updated the datasheet to reflect this 100% predicted cross reactivity.

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

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.