Anti-CD4 antibody [SP35]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [SP35] (AB213215)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human tonsil tissue sections labeling CD4 with ab213215 at 1/50 dilution (3.1 μg/ml). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 10mins. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Hematoxylin was used as a counterstain. Positive staining on the human tonsil, performed on a Leica Biosystems BOND™ RX instrument.
The section was incubated with ab213215 for 30 mins at room temperature.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [SP35] (AB213215)

Immunohistochemical analysis of formalin-fixed, paraffin-embedded human tonsil tissue labeling CD4 with ab213215 at 1/50 dilution.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [SP35] (AB213215)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human Hodgkin lymphoma tissue sections labeling CD4 with ab213215 at 1/50 dilution (3.1 μg/ml). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 10mins. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Hematoxylin was used as a counterstain. Positive staining on the human Hodgkin's lymphoma, performed on a Leica Biosystems BOND™ RX instrument.
The section was incubated with ab213215 for 30 mins at room temperature.

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-CD4 antibody [SP35] (AB213215)

Flow cytometric analysis of Jurkat (Human T cell leukemia cell line from peripheral blood) cells labeling CD4 with ab213215 at 1/100 dilution (green), compared with a rabbit IgG negative control (blue).

• Flow Cyt

Unknown

Flow Cytometry - Anti-CD4 antibody [SP35] (AB213215)

Flow cytometry analysis of THP-1 (human acute monocytic leukemia) labeling CD4 with purified ab213215 at 1/20 dilution (7.75 μg/ml) (red).

Goat anti rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/2000 dilution was used as a secondary antibody. Isotype control - Rabbit monoclonal IgG (ab172730) (Black). Unlableled control - Unlabeled cells (blue).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [SP35] (AB213215)

Immunohistochemical analysis of formalin-fixed, paraffin-embedded human tonsil tissue labeling CD4 with ab213215 at 1/50 dilution.

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-CD4 antibody [SP35] (AB213215)

This data was developed using ab238798 , the same antibody clone in a different buffer formulation. Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Human tonsil tissue labeling CD8 alpha with ab245118 at 1/500 dilution, CD4 with ab238798 at 1/500, and CD19 with ab237772 at 1/5000 dilution. Panel A : merged staining of anti-CD8 alpha (magenta; Opal™690), anti-CD4 (green; Opal™520) and anti-CD19 (red; Opal™570) on human tonsil. Panel B : anti-CD8 alpha stained on cytotoxic T cells. Panel C : anti-CD4 stained on T helper cells. Panel D : anti-CD19 stained on B cells. Sections  were treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins before antibody incubation. The section was incubated in three rounds of staining : in the order of abab245118 for 30 mins, then ab238798 and ab237772 for 10 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. DAPI was used as a nuclear counterstain. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [SP35] (AB213215)

Tissue Microarrays stained for "Anti-CD4 antibody [SP35]" using "ab213215"in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negative (cross mark) staining per sample type tested. The sections were pre-treated using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 10 minutes. The sections were incubated with ab213215 for 30 mins at room temperature followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-CD4 antibody [SP35] (AB213215)

This data was developed using ab238798 , the same antibody clone in a different buffer formulation. Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Human spleen tissue labeling CD8 alpha with ab245118 at 1/500 dilution, CD4 with ab238798 at 1/500, and CD19 with ab237772 at 1/5000 dilution. Panel A : merged staining of anti-CD8 alpha (magenta; Opal™690), anti-CD4 (green; Opal™520) and anti-CD19 (red; Opal™570) on human spleen. Panel B : anti-CD8 alpha stained on cytotoxic T cells. Panel C : anti-CD4 stained on T helper cells. Panel D : anti-CD19 stained on B cells. Sections  were treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins before antibody incubation. The section was incubated in three rounds of staining : in the order of abab245118 for 30 mins, then ab238798 and ab237772 for 10 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. DAPI was used as a nuclear counterstain. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [SP35] (AB213215)

Immunohistochemical analysis of formalin fixed paraffin embedded human tonsil labelling CD4 with ab213215 at a dilution of 1/50. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins.

Anti-CD4 antibody [SP35] was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-CD4 antibody [SP35] (AB213215)

Immunocytochemistry/ Immunofluorescence analysis of human PBMC (human primary peripheral blood mononuclear cell) labeling CD4 with ab238798 at 1/100 dilution followed by ab150081 at 1/1000 dilution (green). The counterstain was Anti-human CD3 mouse monoclonal antibody (Alexa Fluor® 64) at 1/200 dilution observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue). Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% Tween-20. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).