CD4 Antibody (YA537)

MedchemExpress Validation
Immunohistochemical analysis of paraffin-embedded human Colon cancer‌ tissue using CD4 antibody was performed. The section was pretreated using high-temperature and high-pressure mediated EDTA antigen retrieval buffer (pH 9.0), for 5 minutes. The tissues were incubated with primary antibody (HY-P80055, 1:400 dilution) at room temperature for 60 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 Tonsil tissue using CD4 antibody was performed. The section was pretreated using high-temperature and high-pressure mediated EDTA antigen retrieval buffer (pH 9.0), for 5 minutes. The tissues were incubated with primary antibody (HY-P80055, 1:400 dilution) at room temperature for 60 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
Tyramide signaling amplification based immunofluorescence was performed on paraffin-embedded human Liver Cancer tissue using CD4 antibody. Antigen retrieval was performed in EDTA buffer pH 9.0 (95 °C, 20 min) followed by cooling to RT. Then incubated with primary antibody (HY-P80055, 1:400 dilution) at room temperature for 60 minutes and HRP conjugated secondary antibody for 10 minutes. Fluorescence was then developed with TSA520. The tissues were counterstained with DAPI and mounted with Anti-fade mounting medium.

MedchemExpress Validation
Tyramide signaling amplification based immunofluorescence was performed on paraffin-embedded human Esophageal Carcinoma tissue using CD4 antibody. Antigen retrieval was performed in EDTA buffer pH 9.0 (95 °C, 20 min) followed by cooling to RT. Then incubated with primary antibody (HY-P80055, 1:400 dilution) at room temperature for 60 minutes and HRP conjugated secondary antibody for 10 minutes. Fluorescence was then developed with TSA520. The tissues were counterstained with DAPI and mounted with Anti-fade mounting medium.

MedchemExpress Validation
Tyramide signaling amplification based immunofluorescence was performed on paraffin-embedded human Prostate Cancer tissue using CD4 antibody. Antigen retrieval was performed in EDTA buffer pH 9.0 (95 °C, 20 min) followed by cooling to RT. Then incubated with primary antibody (HY-P80055, 1:400 dilution) at room temperature for 60 minutes and HRP conjugated secondary antibody for 10 minutes. Fluorescence was then developed with TSA520. The tissues were counterstained with DAPI and mounted with Anti-fade mounting medium.

MedChemExpress Validation
Flow cytometric analysis of 1X10⁶ THP-1 cells labeling CD4 Antibody (HY-P80055, red). Cells were stained with the primary antibody at 1/500 dilution for an hour at 4℃. AF488-conjugated Goat Anti-Rabbit IgG H&L (HY-P8002) was used as the secondary antibody at 1/1,000 dilution for 30 minutes at 4℃. Rabbit IgG Isotype Control (HY-P80879, 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 CD4 Antibody (HY-P80055, red). Cells were stained with the primary antibody at 1/500 dilution for an hour at 4℃. AF488-conjugated Goat Anti-Rabbit IgG H&L (HY-P8002) was used as the secondary antibody at 1/1,000 dilution for 30 minutes at 4℃. Rabbit IgG Isotype Control (HY-P80879, blue) was used as the isotype control, cells without incubation with primary antibody were used as the unlabeled control (black).

MedchemExpress Validation
Immunocytochemistry analysis of Hela cells labeling CD4 with CD4 Antibody (HY-P80055)at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with CD4 Antibody (HY-P80055) at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure 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 HepG2 cells labeling CD4 with CD4 Antibody (HY-P80055) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with CD4 Antibody (HY-P80055) at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure 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).