Human B7-H3 Antibody

B7-H3 in Human Lung Tumor via seqIF™staining on COMET™ B7-H3 was detected in immersion fixed paraffin-embedded sections of human Lung Tumor using Rabbit Anti-Human B7-H3, Monoclonal Antibody (Catalog #MAB11721) at 1ug/mL at 37 ° Celsius for 4 minutes. Before incubation with the primary antibody, tissue underwent an all-in-one dewaxing and antigen retrieval preprocessing using PreTreatment Module (PT Module) and Dewax and HIER Buffer H (pH 9; Epredia Catalog # TA-999-DHBH). Tissue was stained using the Alexa Fluor™ Plus 647 Goat anti-Rabbit IgG Secondary Antibody at 1:200 at 37 ° Celsius for 2 minutes. (Yellow; Lunaphore Catalog # DR647RB) and counterstained with DAPI (blue; Lunaphore Catalog # DR100). Specific staining was localized to the membrane. Protocol available in COMET™ Panel Builder.

Detection of B7‑H3 in Human Liver Cancer. B7‑H3 was detected in immersion fixed paraffin-embedded sections of human liver cancer using Rabbit Anti-Human B7‑H3 Monoclonal Antibody (Catalog # MAB11721) at 0.5 µg/ml for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC003) or the HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using VisUCyte Antigen Retrieval Reagent-Basic (Catalog # VCTS021). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to the membrane. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Detection of B7‑H3 in Human Lung Cancer. B7‑H3 was detected in immersion fixed paraffin-embedded sections of human lung cancer using Rabbit Anti-Human B7‑H3 Monoclonal Antibody (Catalog # MAB11721) at 0.5 µg/ml for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC003) or the HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using VisUCyte Antigen Retrieval Reagent-Basic (Catalog # VCTS021). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to the membrane. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Detection of Human B7‑H3 by Western Blot. Western Blot shows lysates of JAR human choriocarcinoma cell line and LNCaP human prostate cancer cell line. PVDF membrane was probed with 0.25 µg/ml of Rabbit Anti-Human B7‑H3 Monoclonal Antibody (Catalog # MAB11721) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for B7‑H3 at approximately 90-110 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Detection of Human B7‑H3 by Simple Western^TM. Left: Simple Western lane view shows lysates of LNCaP human prostate cancer cell line, loaded at 0.1 mg/ml. A specific band was detected for B7‑H3 at approximately 149 kDa (as indicated) using both 2 µg/ml and 10 µg/ml of Rabbit Anti-Human B7‑H3 Monoclonal Antibody (Catalog # MAB11721) followed by HRP-conjugated Goat Anti-Rabbit Secondary Antibody (Catalog # 042-206). This experiment was conducted under reducing conditions and using the 12-230kDa separation system. Right: Simple Western electropherogram showing the same Rabbit Anti-Human B7‑H3 Monoclonal Antibody (Catalog # MAB11721) tested at 2 µg/ml (blue line) and 10 µg/ml (green line) in the LNCaP human prostate cancer cell line.