Anti-Hepatitis B Virus Core Antigen antibody [14E11] (ab8638)

We unfortunately do not sell starter packs so I am sorry we cannot send the same. These products are stored in standard amount and the information is available on the datasheets.

Thank for your inquiry. I would like to reassure you that the antibody is the same as previously. There is no batch to batch variability of the immunogen sequence in monoclonal antibodies. This a monoclonal antibody was known to bind to the HBV Core Antigen which corresponds to the area of amino acid residues 130-140. More recently we discovered that the epitope had been mapped and the literature indicates that this clone (14E11) recognizes the amino acids 135-141 which corresponds to the following sequence: PNAPILS. I hope this information is helpful. Please do not hesitate to contact me again with any further questions.

1. The epitope is 135-141 (PNAPILS) 2. This depends on how much virus or core antigen is in the cells. You can load different amounts in parallel. For example 5ug (microgram) and 20ug. 3. Example version of Laemmli 2X buffer 4% SDS 10% 2-mercaptoehtanol 20% glycerol 0.004% bromophenol blue 0.125 M Tris HCl Check the pH and bring it to pH 6.8. Our online protocols are at www.abcam.com/protocols I highly recommend to look at the publications: PubMed: 19327810 and 16378961.

Thank you for your enquiry. Further to discussions with the lab I have determined that the approach that you are using (n)on-reducing gel at 12%) is not the best system to isolate monomeric units of HBcAg core particles. To dissolve HBcAg core particles to monomers, the sample should be dissolved in the Laemmli buffer with 5%SDS, 5% Mercaptoethanol (added fresh each time), and boiled for 10 min on a water bath. Should you continue to get what you expect to be polymers of this protein please do not hesitate to get back in touch with me.

Thank you for your enquiry. I am sorry to hear that you have been having difficulties with this antibody. I am surprised by the size of the band that you have been detecting. I am in correspondence with the source of this antibody to determine the expected size of the band using this antibody. At this stage I would like to recommend that you transfer the protein lysate that you have prepared and cut the blot using a scalpel so that only proteins of a molecular weight less than 100KDa are immunoblotted and the antibody is concentrated over proteins of appropriate size. I would like to recommend that an overnight incubation is performed using 3% BSA as a blocking agent. This approach should significantly increase the chances of detecting the Hepatitis core antigen. I hope this information helps. Should this approach not improve your results please do not hesitate to get back in touch with me.

Ab8637 and ab8639 should recognize the precorerotein in WB but there is no "cross-reaction" as all proteins are distinguishable by mobility on the gel (i.e they will recognise both the precoreprotein and core protein). AB8638 may or may not recognize your protein fragment aa1-150, since its epitope is aa aa35-140 of the core protein, which is really close to the N terminus of the protein you are going to analyze. Ab8637 will not recognize the precorprotein under native conditions, because this protein can not self assemble into particles. I hope this information will help you, please let me know if you need further assistance,

Here is the information that I could obtain regarding ab8639, ab8638, and ab8637. The recommended starting dilutions are 1:1,000 and down in 10-fold increments. As for detection, any known detection method would work. Once again, those mabs had being characterized in great detail in the following paper: Bichko et al. Epitopes recognized by antibodies to denatured core protein of hepatitis B virus. Mol Immunol. 1993 Feb;30(3):221-31 If you have any more questions, just let me know.

I have received the following information regarding ab8255 and ab2045, but will have to get back to you about the other antibodies. The originator has said that they tested the antibodies as pairs in internal ELISA. Possible pairs are H6F5 (ab2045) - H3A4 (ab8255), and H3A4 - H6F5. They have used coating concentration 5 - 10 ug/ml and conjugate concentration 0.2 -1 ug/ml. However, the concentrations need to be optimized separately for every application.

Thank you for your enquiry. I was able to obtain the following information regarding the use of these antibodies in ELISA. Ab8255 and Ab2045: These antibodies have been tested in ELISA with the native antigen. The antibodies can be used as a pair in sandwich ELISA. Both clones can work as capture and as detection antibodies. Ab8638, Ab8637, and Ab8639: These antibodies work well with a recombinant or natural HBcAg adsorbed on a plate, or in sandwich ELISA. Also, they work well with the synthetic HBcAg peptides. Below is one reference: Bichko et al. Epitopes recognized by antibodies to denatured core protein of hepatitis B virus. Mol Immunol. 1993 Feb;30(3):221-31

Thank you for your reply. The only data we have regarding this is that it is an ayw variant as described in: Bichko et. al. Subtype ayw variant of hepatitis B virus. DNA primary structure analysis. FEBS Lett. 1985 Jun 3;185(1):208-12.