Anti-Hepatitis B Virus X antigen antibody [3F6-G10] (ab235)

Sorry for the delay in getting back to you. I now have further information to share with you.

I have checked into whether anything is know about the epitope of the anti-Hepatitis B Virus X antigen antibodies ab235, ab2741 and ab39716. I can only provide further information for ab235 additionally to the information provided on the datasheet of the antibodies. The epitope recognised by ab235 is thought to recognise an epitope of the following sequence:

DVLCLRPVGASESR.

I was incorrect in my last email to say that ab235 had not been tested in Western blotting. It have been tested but we have had mixed reports from customers so we would not recommend buying this antibody with that application in mind.

I have also looked into the anti-Hepatitis B Virus Core Antigen antibodies we have and if anything is known of the conformation of the epitope is known. Unfortunately, I could not find any antibodies where this information had been established and most are able to recognise both the denatured and native form of the protein.

I hope this information has been of help. If you have any further questions, please do not hesitate to ask.

Thank you for contacting us and your interest in our products.

The request for a quote for the antibodies mentioned has been passed on to one of my colleagues who will get back to you shortly with this information. In regards to your other questions:

The Rabbit anti-Mouse IgG H&L (HRP) secondary antibody (ab7075) should work well with ab15149, ab2741, ab235 and ab26. You would need a different secondary antibody for all the other antibodies mentioned as they are raised in rabbit. You may wantto consider the Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP), pre-adsorbed (ab97080). Please bear in mind that we have as yet not tested the anti-Hepatitis B Virus X antigen antibody [3F6-G10] (ab235) in Western blotting or immunoprecipitation. This does not mean that it would not work, simply that we have not tested it ourselves as yet and would therefore not he able to guarantee its performance in these applications.

We have some good resources in setting up both immunoprecipitation and Western blotting experiments from the following link:

https://docs.abcam.com/pdf/misc/abcam-protocols-book-2010.pdf

Some further troubleshooting tips can also be found here:

https://www.abcam.com/index.html?pageconfig=resource&rid=11382

If you would like any help with this please do let us know.

I will check if any further information is know about the epitopes recognised by the anti-HBx antibodies mentioned and will get back to you with this information as soon as I have it. I will also look into your final question ifwe have any conformation specific antibodies against HBV core protein (HBc).

As you will be setting up these experiments from scratch, you may be interested in looking at our Western blotting range:

https://www.abcam.com/index.html?pageconfig=resource&rid=14709&source=pagetrap&viapagetrap=optiblot

The link above will take you not only to our Optiblot range of gels which provide superior durability and ease of use compared to many standard gels, but also the PVDF and nitrocellulose membranes and Ecl Detection kits we have available.

If you would like any further information on these products please do let us know.

Thank you for contacting us. The note on the datasheet is from the originator of the 3F6-G10 clone. Production of the clone and testing against the recombinant protein is described in the following paper: Pál J et al. Immunohistochemical assessment and prognostic value of hepatitis B virus X protein in chronic hepatitis and primary hepatocellular carcinomas using anti-HBxAg monoclonal antibody. Pathol Oncol Res 7:178-84 (2001). PubMed: 11692143. The protein is not commercially available, as far as we know. If you are having difficulty with the antibody, please let us know and we will help resolve the problem.

Thank you for your message. Unfortuantley, I am unable to obtain precise information about which subtypes of Hepatitis B virus these antibodies detect. We aim to provide as much information as possible to customers, and I am sorry this is not possible on this occasion. With regards to whether the antibodies detect native or denatured protein, you should always use reducing and denaturing conditions unless stated otherwise on the datasheet. Please also check the information I have provided below for futher details. I did manage to obtain the following information on these antibodies, which I hope you will find helpful: Reference for ab2741. This reference uses the antibody in IP and WB procedures. M. Melegari, et al. Cloning and Characterization of a Novel Hepatitis B Virus x Binding Protein That Inhibits Viral Replication. J. Virology, 72(3): 1737-1743, 1998. http://jvi.asm.org/cgi/content/full/72/3/1737 Hepatitis B Virus X antigen antibody ab39716 was tested only with recombinant antigen in WB (reducing and nonreducing) and for immunoprecipitation: with antigen from HeLa cells transfected with HBX tagged with c-myc. Antibody ab235 reacts with both the native and denatured HBX antigens. For further details of this clone please see the following references: Pal J, Somogyi C, Szmolenszky A A, Szekeres G, Sipos J, Hegedus G, Martzinovits I, Molnar J, Nemeth P. Immunohistochemical assessment and prognostic value of hepatitis B virus X protein in chronic hepatitis and primary hepatocellular carcinomas using anti-HBxAg monoclonal antibody. Pathol Oncol Res. 2001;7(3):178-84. PMID: 11692143 Marczinovits I, Somogyi C, Patthy A, Nemeth P, Molnar J. An alternative purification protocol for producing hepatitis B virus X antigen on a preparative scale in Escherichia coli. J Biotechnol. 1997 Aug 11;56(2):81-8. PMID: 9304871 I hope this information is helpful. Should you have any further questions, please do not hesiate to contact us again.