Anti-GITR antibody [CAL8]

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-GITR antibody [CAL8] (AB237714)

Intracellular flow cytometric analysis of 2% paraformaldehyde-fixed,0.1% Tween-20permeabilized human peripheral blood mononuclear cell (PBMC) treated with 10μg/ml PHA for 48hrs, labeling GITR with ab237714 at 1/500 (right) compared with a rabbit IgG (left). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077), at 1/5000 dilution was used as the secondary antibody. Cells were surface stained with anti-CD25 conjugated to BV421. Then fixed with 2% PFA followed by intracellular staining rabbit IgG (Left) or ab237714 (Right).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GITR antibody [CAL8] (AB237714)

Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling GITR with ab237714 at 1/500 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining on the human tonsil tissue 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

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GITR antibody [CAL8] (AB237714)

Formalin-fixed, paraffin-embedded human tonsil tissue stained for TNFSF18 using ab237714 at 0.5 μg/ml in immunohistochemical analysis.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-GITR antibody [CAL8] (AB237714)

Flow cytometry staining of human peripheral blood mononuclear cells (PBMCs) (top) or PBMCs treated with anti-CD3 and anti-CD28 antibodies (72 hours) (bottom), with ab237714 (right) or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (left). Cells were fixed and permeabilised with BD Cytofix/Cytoperm™ for 20 min. PBMCs were incubated for 30 min at 4°C in 1x PBS containing 10 μg/ml human IgG and 10 % normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody ab237714 or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (1x 106 in 100 μl at 0.2 μg/ml (1/10200)) for 30 min at 4°C . The cells were simultaneously stained with CD8.

The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min at 4°C

Acquisition of >30000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter. Events were gated on viable cells.