Human p62/SQSTM1 Antibody

Detection of Human p62/SQSTM1 by Western Blot. Western blot shows lysates of A172 human glioblastoma cell line and RT-4 human bladder carcinoma cell line. PVDF membrane was probed with 1 µg/mL of Rabbit Anti-Human p62/SQSTM1 Monoclonal Antibody (Catalog # MAB80281) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (HAF008). A specific band was detected for p62/SQSTM1 at approximately 60 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

p62/SQSTM1 in HeLa Human Cell Line. p62/SQSTM1 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line treated with chloroquine (left) or untreated (right) using Rabbit Anti-Human p62/SQSTM1 Monoclonal Antibody (Catalog # MAB80281) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; NL004) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

p62/SQSTM1 in Human Liver. p62/SQSTM1 was detected in immersion fixed paraffin-embedded sections of human liver using Rabbit Anti-Human p62/SQSTM1 Monoclonal Antibody (Catalog # MAB80281) at 10 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte™ HRP Polymer Antibody (VC003). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cell nuclei. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Detection of Human p62/SQSTM1 by Simple Western^TM. Simple Western lane view shows lysates of RT-4 human bladder carcinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for p62/SQSTM1 at approximately 64 kDa (as indicated) using 20 µg/mL of Rabbit Anti-Human p62/SQSTM1 Monoclonal Antibody (Catalog # MAB80281). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Western Blot Shows Human p62/SQSTM1 Specificity by Using Knockout Cell Line. Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and p62/SQSTM1 knockout HeLa cell line (KO). PVDF membrane was probed with 1 µg/mL of Rabbit Anti-Human p62/SQSTM1 Monoclonal Antibody (Catalog # MAB80281) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (HAF008). A specific band was detected for p62/SQSTM1 at approximately 60 kDa (as indicated) in the parental HeLa cell line, but is not detectable in knockout HeLa cell line. GAPDH (2275-PC-100) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Western Blot Shows Human p62/SQSTM1 Specificity Using Knockout Cell Line. Western blot shows lysates of U2OS human osteosarcoma cell line and p62/SQSTM1 knockout U2OS cell line (KO). Nitrocellulose membrane was probed with 0.5 µg/mL of Rabbit Anti-Human p62/SQSTM1 Monoclonal Antibody (Catalog # MAB80281) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody. A specific band was detected for p62/SQSTM1 at approximately 62 kDa (as indicated) in the parental U2OS cell line, but is not detectable in knockout U2OS cell line. The Ponceau stained transfer of the blot is shown. This experiment was conducted under reducing conditions. Image, protocol, and testing courtesy of YCharOS Inc. See ycharos.com for additional details.

Detection of SQSTM1/p62 by Immunoprecipitation. Immunoprecipitation was performed on cell lysate of U2OS human osteosarcoma cell line using 1.0 μg of Mouse Anti-Human SQSTM1/p62 Monoclonal Antibody (Catalog # MAB80281) pre-coupled to protein G or protein A beads. Immunoprecipitated SQSTM1/p62 was detected with a Mouse Anti-SQSTM1/p62 Monoclonal Antibody. The Ponceau stained transfers of each blot are shown. SM=10% starting material; UB=10% unbound fraction; IP=immunoprecipitated. Image, protocol, and testing courtesy of YCharOS Inc. (ycharos.com).