Rabbit anti hbcag

Manufactured by Agilent Technologies

Sourced in Denmark

Rabbit anti-HBcAg is a laboratory reagent used for the detection and analysis of the hepatitis B core antigen (HBcAg) in biological samples. It is a polyclonal antibody produced by immunizing rabbits with HBcAg. This reagent can be used in various immunoassay techniques, such as ELISA, immunoblotting, and immunohistochemistry, to identify the presence and quantify the levels of HBcAg in samples.

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Lab products found in correlation

Complete mini protease inhibitor cocktail, by Roche (1 mentions) Brightvision poly hrp detection system, by Immunologic (1 mentions) Visiomorph, by Visiopharm (1 mentions) Hoechst 33258, by Thermo Fisher Scientific (1 mentions) Fluorescent mounting media, by Agilent Technologies (1 mentions) Anti rabbit horseradish peroxidase conjugated secondary antibody, by Jackson ImmunoResearch (1 mentions) Tsa fluorescein system, by PerkinElmer (1 mentions) Mouse anti ck18, by Agilent Technologies (1 mentions)

Spelling variants of this product

Anti-HBcAg (12 citations) Polyclonal rabbit anti-HBcAg (3 citations) Rabbit anti- HBcAg polyclonal antibody (2 citations) Polyclonal rabbit anti-HBcAg primary antiserum (2 citations) Anti-TM (1 citations)

4 protocols using rabbit anti hbcag

Western Blot Analysis of HBcAg

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Cells were treated with different concentrations of compounds for six days, washed with cold PBS, and lysed with radio immunoprecipitation assay (RIPA) lysis buffer supplemented with complete mini protease inhibitor cocktail (Roche) at 4 °C for 1 h. Cell lysates were subjected to 15% SDS-PAGE and transferred to poly-vinylidene difluoride (PVDF) membranes. After incubation with rabbit anti-HBcAg (Dako) at 4 °C overnight, each blot was probed with horseradish peroxidase-conjugated secondary antibody. Immunoreactive signals were detected with an enhanced chemiluminescence substrate (Thermo) using an AlphaEaseH FC Imaging System (Alpha Innotech Corporation).

Liu C., Cai D., Zhang L., Tang W., Yan R., Guo H, & Chen X. (2016). Identification of hydrolyzable tannins (punicalagin, punicalin and geraniin) as novel inhibitors of hepatitis B virus covalently closed circular DNA. Antiviral research, 134, 97-107.

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Quantitative Assessment of HBcAg in Mouse Liver

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Two sections separated by approximately 200 μm of formalin-fixed and paraffin-embedded mouse liver were deparaffinized and subjected to heat-induced antigen retrieval in Tris-EDTA glucose (TEG) buffer (pH 9). Following blockage of endogenous peroxidase activity, the sections were blocked in 10% normal serum and incubated with rabbit anti-HBcAg (Dako). The primary antibody was detected and amplified using Brightvision Poly-HRP detection system (Immunologic) and visualized with diaminobenzidine as chromogen. Finally, sections are counterstained in hematoxylin, coverslipped, and digitized using a 20× objective. Quantitative assessment of HBcAg immunoreactivity was estimated as total counts of positive cellular profiles per area of the liver sections. Profile counting is done by image analysis using Visiomorph (Visiopharm).

Javanbakht H., Mueller H., Walther J., Zhou X., Lopez A., Pattupara T., Blaising J., Pedersen L., Albæk N., Jackerott M., Shi T., Ploix C., Driessen W., Persson R., Ravn J., Young J.A, & Ottosen S. (2018). Liver-Targeted Anti-HBV Single-Stranded Oligonucleotides with Locked Nucleic Acid Potently Reduce HBV Gene Expression In Vivo. Molecular Therapy. Nucleic Acids, 11, 441-454.

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HBV Infection Visualization in HepG2-NTCP Cells

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HepG2-NTCP cells were infected by HBV for 3 days, followed by fixation with 4% paraformaldehyde for 20 min and permeabilized in 0.5% Triton X-100 1×PBS solution for 1 h at room temperature. Subsequently, cells were blocked with IFA blocking buffer (10% FBS plus 2% bovine serum albumin in 1×PBS) for 1 h at room temperature and incubated with rabbit anti-HBcAg (cat# B0586, Dako) at 4°C for overnight. After washing by 1×PBS and a second blocking for 30 min, cells were incubated with Alexa Fluor 488 dye-conjugated goat anti-rabbit secondary antibody (cat# A11070, Invitrogen), and cell nuclei were counterstained with DAPI (4’,6-diamidino-2-phenylindole) for 30 min at room temperature. Both primary and secondary antibodies were diluted in IFA blocking buffer. The cells were washed with 1×PBS and subjected to Olympus FV1000 microscopy analysis under 20× objective lens. Images were analyzed using Image J software.

Yu X., Long Q., Shen S., Liu Z., Chandran J., Zhang J., Ding H., Zhang H., Cai D., Kim E.S., Huang Y, & Guo H. (2023). Screening of an epigenetic compound library identifies BRD4 as a potential antiviral target for hepatitis B virus covalently closed circular DNA transcription. Antiviral research, 211, 105552.

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Immunohistochemical Staining of Chimeric Mouse Livers

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Cryostat sections of chimeric mouse livers were stained as previously described (Lutgehetmann et al., 2012 (link)). Briefly, sections were fixed with acetone and incubated with mouse anti-CK18 (1:400, Dako, Glostrup, Denmark), rabbit anti-HBcAg (1:2000, Dako), mouse HLA-ABC (1:50, Antibodies-online, Aachen, Germany), and human anti-Delta (anti-HDAg-positive human serum, 1:8,000). Specific signals were visualized with Alexa 488-, 555-, or 633-labeled secondary antibodies (Invitrogen, Darmstadt, Germany). To enhance the HBcAg staining an anti-rabbit horseradish-peroxidase conjugated secondary antibody (Jackson Immunoresearch, Suffolk, United Kingdom) and the TSA Fluorescein System (Perkin Elmer, Jügesheim, Germany) were used. Nuclear staining was achieved by Hoechst 33258 (1:20,000 diluted, Invitrogen, Waltham, MA, United States). Stained sections were then mounted with fluorescent mounting media (Dako) and analyzed with the fluorescence microscope BZ8710 (Keyence, Osaka, Japan) using the same settings for the different experimental groups. The percentages of HDAg-positive human hepatocytes were estimated as previously described (Lutgehetmann et al., 2012 (link)) and by using 2–5 visual fields (displaying an average of 500 human hepatocytes) per mouse liver.

Giersch K., Hermanussen L., Volz T., Volmari A., Allweiss L., Sureau C., Casey J., Huang J., Fischer N., Lütgehetmann M, & Dandri M. (2021). Strong Replication Interference Between Hepatitis Delta Viruses in Human Liver Chimeric Mice. Frontiers in Microbiology, 12, 671466.

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