CD8 alpha Antibody(YA6867)

MedChemExpress Validation
Western blot analysis was performed on protein extracts (25 μg) from Mouse spleen (lane 2), Mouse thymus (lane 3) and Mouse lung (lane 4) using CD8 alpha 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. The primary antibody (1:1000) and loading control antibody GAPDH Antibody (HRP) (HY-P80954A) (1:5000) 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-rabbit/mouse IgG secondary antibody (HY-P8001/HY-P8004) (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 mouse spleen tissue using CD8 alpha Antibody(YA6867). The section was pre-treated using heat mediated antigen retrieval with Tris/EDTA buffer (pH 9.0) for 20 minutes. The tissues were probed with the primary antibody (HY-P87176, 1/200) 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 neutral balsam.

MedChemExpress Validation
Immunohistochemical analysis of paraffin-embedded mouse jejunum tissue using CD8 alpha Antibody(YA6867). The section was pre-treated using heat mediated antigen retrieval with Tris/EDTA buffer (pH 9.0) for 20 minutes. The tissues were probed with the primary antibody (HY-P87176, 1/200) 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 neutral balsam.

MedChemExpress Validation
Immunohistochemical analysis of paraffin-embedded mouse liver tissue using CD8 alpha Antibody(YA6867). The section was pre-treated using heat mediated antigen retrieval with Tris/EDTA buffer (pH 9.0) for 20 minutes. The tissues were probed with the primary antibody (HY-P87176, 1/200) 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 neutral balsam.

MedChemExpress Validation
Immunohistochemical analysis of paraffin-embedded mouse lung tissue using CD8 alpha Antibody(YA6867). The section was pre-treated using heat mediated antigen retrieval with Tris/EDTA buffer (pH 9.0) for 20 minutes. The tissues were probed with the primary antibody (HY-P87176, 1/200) 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 neutral balsam.

MedChemExpress Validation
Immunohistochemical analysis of paraffin-embedded mouse thymus tissue using CD8 alpha Antibody(YA6867). The section was pre-treated using heat mediated antigen retrieval with Tris/EDTA buffer (pH 9.0) for 20 minutes. The tissues were probed with the primary antibody (HY-P87176, 1/200) 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 neutral balsam.

MedChemExpress Validation
Immunocytochemistry analysis of MCF-7 cells labeling CD8 alpha with CD8 alpha Antibody (HY-P87176) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with quick block buffer for 10 minutes at room temperature. Cells were then incubated with CD8 alpha Antibody (HY-P87176) at 1/100 dilution in quick block buffer overnight at 4 ℃. AF488-conjugated Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).

MedChemExpress Validation
Immunocytochemistry analysis of U2OS cells labeling CD8 alpha with CD8 alpha Antibody (HY-P87176) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with quick block buffer for 10 minutes at room temperature. Cells were then incubated with CD8 alpha Antibody (HY-P87176) at 1/100 dilution in quick block buffer overnight at 4 ℃. AF488-conjugated Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).