Anti-CD4 antibody [CAL4]

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-CD4 antibody [CAL4] (AB237722)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse spleen tissue labeling CD4 with ab237722 at 1/100 dilution (green), followed by ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at a 1/1,000 dilution. Positive staining on T cell surface (PMID : 26994219) is observed. Counterstained with DAPI (blue). Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is ab150077 AlexaFluor^®488 Goat anti-Rabbit used at a 1/1,000 dilution. Heat mediated antigen retrieval using sodium citrate buffer (10 mM citrate pH 6.0 and 0.05% Tween-20).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [CAL4] (AB237722)

Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling CD4 with ab237722 at 1/4000 dilution, followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on rat spleen. The section was incubated with ab237722 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin. Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-CD4 antibody [CAL4] (AB237722)

Intracellular flow cytometric analysis of 2% paraformaldehyde-fixed, 0.1% Tween-20 permeabilized mouse splenocytes labeling CD4 with ab237722 at 1/40 (right panel) compared with a Rabbit monoclonal IgG (ab172730) (left panel). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077), at 1/2000 dilution was used as the secondary antibody. Cells were surface stained with anti-CD4 conjugated to Alexa Fluor^® 647. Then fixed with 2% PFA for 10min and intracellularly stained with rabbit IgG (Left) or ab237722 (Right). They recognize the same cell population.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [CAL4] (AB237722)

Formalin-fixed, paraffin-embedded mouse spleen tissue stained for CD4 using ab237722 at 0.5 μg/ml in immunohistochemical analysis.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [CAL4] (AB237722)

Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling CD4 with ab237722 at 1/4000 dilution, followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse spleen. The section was incubated with ab237722 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin. Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

• IP

Lab

Immunoprecipitation - Anti-CD4 antibody [CAL4] (AB237722)

CD4 was immunoprecipitated from 0.35 mg mouse thymus lysate with ab237722 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab237722 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used at 1/5000 dilution.
Lane 1 : Mouse thymus lysate 10 μg (Input).
Lane 2 : ab237722 IP in mouse thymus lysate .
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab237722 in mouse thymus lysate .
Blocking/Dilution buffer : 5% NFDM/TBST.
Exposure time : 30 seconds.

102kDa band represents CD4 dimer based on literature. (PMID : 20739067).

All lanes:

Immunoprecipitation - Anti-CD4 antibody [CAL4] (ab237722)

Predicted band size: 51 kDa

false

• WB

Lab

Western blot - Anti-CD4 antibody [CAL4] (AB237722)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-CD4 antibody [CAL4] (ab237722) at 1/1000 dilution

Lane 1:

Mouse thymus lysate at 20 µg

Lane 2:

Mouse spleen lysate at 20 µg

Lane 3:

Mouse lymph node lysate at 20 µg

Lane 4:

Rat thymus lysate at 20 µg

Lane 5:

Rat spleen lysate at 20 µg

Lane 6:

Rat lymph node lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 51 kDa

false