CD276/B7H3 Antibody (YA4334)

MedChemExpress Validation
Western blot analysis was performed on protein extracts (30 μg) from HepG2 (lane 2) and Caco-2 (lane 3) using CD276/B7H3 antibody. Proteins were transferred onto a 0.45 μm PVDF membrane using the Trans-Blot^® Turbo^™ system for 13 min. The membrane was then blocked with 5% nonfat milk in TBST (HY-K1025) for 1 h at room temperature. Thhe primary antibody (1:1000) and loading control antibody GAPDH Antibody (HRP) (HY-P80954A) (1:2500) were diluted in 5% nonfat milk in TBST and incubated with the membrane overnight at 4°C. After washing, the membrane of primary antibody was incubated with HRP-conjugated goat anti-mouse IgG (H&L) secondary antibody (HY-P8001) (1:5000) diluted in 5% nonfat milk in TBST for 1 h at room temperature. Protein bands were visualized using an Ultra High Sensitivity ECL detection kit (HY-K1005).

MedChemExpress Validation
Immunohistochemical analysis of paraffin-embedded human cervix tissue using CD276/B7H3 Antibody (HY-P84637, 1/500). The section was pretreated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 8 minutes. The tissues were blocked with quick block buffer for 0.5 hours at room temperature, washed with PBS and PBST, and then incubated with the primary antibody overnight at 4℃. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

MedChemExpress Validation
Immunohistochemical analysis of paraffin-embedded human gallbladder tissue using CD276/B7H3 Antibody (HY-P84637, 1/500). The section was pretreated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 8 minutes. The tissues were blocked with quick block buffer for 0.5 hours at room temperature, washed with PBS and PBST, and then incubated with the primary antibody overnight at 4℃. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

MedChemExpress Validation
Immunohistochemical analysis of paraffin-embedded human tonsil tissue using CD276/B7H3 Antibody (HY-P84637, 1/500). The section was pretreated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 8 minutes. The tissues were blocked with quick block buffer for 0.5 hours at room temperature, washed with PBS and PBST, and then incubated with the primary antibody overnight at 4℃. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

MedChemExpress Validation
Immunohistochemical analysis of paraffin-embedded human thyroid gland tissue using CD276/B7H3 Antibody (HY-P84637, 1/500). The section was pretreated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 8 minutes. The tissues were blocked with quick block buffer for 0.5 hours at room temperature, washed with PBS and PBST, and then incubated with the primary antibody overnight at 4℃. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

MedChemExpress Validation
Immunohistochemical analysis of paraffin-embedded human adrenal gland tissue using CD276/B7H3 Antibody (HY-P84637, 1/500). The section was pretreated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 8 minutes. The tissues were blocked with quick block buffer for 0.5 hours at room temperature, washed with PBS and PBST, and then incubated with the primary antibody overnight at 4℃. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

MedChemExpress Validation
Immunohistochemical analysis of paraffin-embedded human endometrium tissue using CD276/B7H3 Antibody (HY-P84637, 1/500). The section was pretreated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 8 minutes. The tissues were blocked with quick block buffer for 0.5 hours at room temperature, washed with PBS and PBST, and then incubated with the primary antibody overnight at 4℃. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

MedChemExpress Validation
Immunohistochemical analysis of paraffin-embedded human Glioma‌ tissue using CD276 theta antibody was performed. The section was pretreated using high-temperature mediated EDTA antigen retrieval buffer (pH 9.0), for 20 minutes. The tissues were incubated with primary antibody (HY-P84637, 1:100 dilution) at room temperature for 20 minutes. Detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. The tissues were counterstained with hematoxylin and mounted with neutral balsam mounting medium.

MedChemExpress Validation
Immunohistochemical analysis of paraffin-embedded human Liver cyst tissue using CD276 theta antibody was performed. The section was pretreated using high-temperature mediated EDTA antigen retrieval buffer (pH 9.0), for 20 minutes. The tissues were incubated with primary antibody (HY-P84637, 1:100 dilution) at room temperature for 20 minutes. Detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. The tissues were counterstained with hematoxylin and mounted with neutral balsam mounting medium.

MedChemExpress Validation
Immunohistochemical analysis of paraffin-embedded human Kimura's disease tissue using CD276 theta antibody was performed. The section was pretreated using high-temperature mediated EDTA antigen retrieval buffer (pH 9.0), for 20 minutes. The tissues were incubated with primary antibody (HY-P84637, 1:100 dilution) at room temperature for 20 minutes. Detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. The tissues were counterstained with hematoxylin and mounted with neutral balsam mounting medium.

MedChemExpress Validation
Immunohistochemical analysis of paraffin-embedded human Mucoepidermoid carcinoma tissue using CD276 theta antibody was performed. The section was pretreated using high-temperature mediated EDTA antigen retrieval buffer (pH 9.0), for 20 minutes. The tissues were incubated with primary antibody (HY-P84637, 1:100 dilution) at room temperature for 20 minutes. Detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. The tissues were counterstained with hematoxylin and mounted with neutral balsam mounting medium.

MedChemExpress Validation
Immunohistochemical analysis of paraffin-embedded human solitary fibrous tumor tissue using CD276 theta antibody was performed. The section was pretreated using high-temperature mediated EDTA antigen retrieval buffer (pH 9.0), for 20 minutes. The tissues were incubated with primary antibody (HY-P84637, 1:100 dilution) at room temperature for 20 minutes. Detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. The tissues were counterstained with hematoxylin and mounted with neutral balsam mounting medium.

MedChemExpress Validation
Immunohistochemical analysis of paraffin-embedded human cervical squamous cell carcinoma tissue using CD276 theta antibody was performed. The section was pretreated using high-temperature mediated EDTA antigen retrieval buffer (pH 9.0), for 20 minutes. The tissues were incubated with primary antibody (HY-P84637, 1:100 dilution) at room temperature for 20 minutes. Detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. The tissues were counterstained with hematoxylin and mounted with neutral balsam mounting medium.

MedChemExpress Validation
Flow cytometric analysis of 1X10⁶ THP-1 cells labeling CD276/B7H3 Antibody (HY-P84637, red). Cells were stained with the primary antibody at 1/200 dilution for an hour at 4℃. AF488-conjugated Goat Anti-Mouse IgG H&L (HY-P8005) was used as the secondary antibody at 1/1,000 dilution for 30 minutes at 4℃. Mouse IgG Isotype Control (HY-P80757, blue) was used as the isotype control, cells without incubation with primary antibody were used as the unlabeled control (black).

MedChemExpress Validation
Flow cytometric analysis of 1X10⁶ THP-1 cells labeling CD276/B7H3 Antibody (HY-P84637, red). Cells were stained with the primary antibody at 1/200 dilution for an hour at 4℃. AF488-conjugated Goat Anti-Mouse IgG H&L (HY-P8005) was used as the secondary antibody at 1/1,000 dilution for 30 minutes at 4℃. Mouse IgG Isotype Control (HY-P80757, blue) was used as the isotype control, cells without incubation with primary antibody were used as the unlabeled control (black).