Anti-GITR antibody [CAL61]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GITR antibody [CAL61] (AB237725)

Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling GITR with ab237725 at 1/500 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining on the mouse spleen is observed. Counterstained with hematoxylin. Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GITR antibody [CAL61] (AB237725)

Formalin-fixed, paraffin-embedded mouse spleen tissue stained for GITR using ab237725 at 0.125 μg/ml dilution in immunohistochemical analysis.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GITR antibody [CAL61] (AB237725)

Immunohistochemical analysis of paraffin-embedded mouse lung tissue labeling GITR with ab237725 at 1/500 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining on the mouse lung is observed. Counterstained with hematoxylin. Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-GITR antibody [CAL61] (AB237725)

Intracellular flow cytometric analysis of 2% paraformaldehyde-fixed,0.1% Tween-20permeabilized mouse splenocytes treated with 2.5μg/ml concanavalin A for 3 days labeling GITR with ab237725 at 1/500 (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-CD25 conjugated to BV421. Then fixed with 2% PFA followed by intracellularly stained with rabbit IgG (Left) or ab237725 (Right).

• WB

Lab

Western blot - Anti-GITR antibody [CAL61] (AB237725)

33 kDa band represents GITR isoform B based on Uniprot and literature. (PMID : 10836847).

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-GITR antibody [CAL61] (ab237725) at 1/1000 dilution

Lane 1:

Untreated mouse splenocytes whole cell lysate at 20 µg

Lane 2:

Mouse splenocytes treated with 2.5μg/ml concanavalin A for 3 days whole cell 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: 26 kDa

false

Exposure time: 3min