Human Cathepsin B Antibody

Detection of Human Cathepsin B by Western Blot. Western blot shows lysates of HepG2 human hepatocellular carcinoma cell line. PVDF membrane was probed with 0.25 µg/mL of Goat Anti-Human Cathepsin B Antigen Affinity-purified Polyclonal Antibody (Catalog # AF953) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). A specific band was detected for Cathepsin B at approximately 25-30 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Cathepsin B in Human Brain. Cathepsin B was detected in immersion fixed paraffin-embedded sections of human brain (cortex) using Goat Anti-Human Cathepsin B Antigen Affinity-purified Polyclonal Antibody (Catalog # AF953) at 10 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; CTS008) and counterstained with hematoxylin (blue). Lower panel shows a lack of labeling if primary antibodies are omitted and tissue is stained only with secondary antibody followed by incubation with detection reagents. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Detection of Human Cathepsin B by Simple Western^TM. Simple Western lane view shows lysates of HepG2 human hepatocellular carcinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for Cathepsin B at approximately 34 kDa (as indicated) using 2.5 µg/mL of Goat Anti-Human Cathepsin B Antigen Affinity-purified Polyclonal Antibody (Catalog # AF953) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Western Blot Shows Human Cathepsin B Specificity by Using Knockout Cell Line. Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and Cathepsin B knockout HeLa cell line (KO). PVDF membrane was probed with 0.25 µg/mL of Goat Anti-Human Cathepsin B Antigen Affinity-purified Polyclonal Antibody (Catalog # AF953) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF017). Specific bands were detected for Cathepsin B at approximately 25-45 kDa (as indicated) in the parental HeLa cell line, but is not detectable in knockout HeLa cell line. GAPDH (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1. New adjunct appears with knockout cell line.

Detection of Cathepsin B in U‑87 (Positive) & Daudi (Negative). Cathepsin B was detected in immersion fixed U‑87 MG human glioblastoma/astrocytoma cell line (Positive) & absent in Daudi human Burkitt's lymphoma cell line (Negative) using Goat Anti-Human Cathepsin B Antigen Affinity-purified Polyclonal Antibody (Catalog # AF953) at 5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cell surface. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Detection of Human Cathepsin B by Western Blot Legumain, cathepsin B and L expressions in cytosolic and membrane fractions of myotubes.Myotubes were treated with (+) or without (−) 30 µM simvastatin for 48 h before subcellular fractionation was performed. One representative immunoblot of legumain, cathepsin B and L in the cytosolic and membrane fractions is shown. All lanes were loaded with equal amount of total proteins and probed with antibodies as indicated. LAMP-2 and alpha -tubulin are shown as cell compartment controls (n = 3). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/24416446), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human Cathepsin B by Western Blot CTSS inhibition does not impair lysosomal activities.(a) OEC-M1 cells were pretreated with vesicle or 100 nM BAF for 1 h and then incubated with 20 μM 6r for 1 h further. Lysosomal proteolytic activities were determined using BODIPY–BSA and quantified through flow cytometry. (b) After 48 h of siRNA knockdown of CTSS and CTSB, the relative expression of CTSS and CTSB were determined through Western blotting (right panel). Lysosomal proteolytic activities were determined using BODIPY–BSA and quantified through flow cytometry (right panel). Data represent the mean ± SD of three independent experiments. Differences were found to be statistically significant at ***P < 0.001. Image collected and cropped by CiteAb from the following open publication (https://www.nature.com/articles/srep29256), licensed under a CC-BY license. Not internally tested by R&D Systems.