Recombinant 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.

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 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).

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.

Foreskin epidermis at 10× magnification stained with CCR5 (pseudocolored green), CD4 (pseudocolored red; ab213215) and SYTOX Orange (pseudocolored blue) for nuclear identification. All fields from ∼26.5 mm² sections were used for analysis.

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.

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).

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

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

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.