Anti-LAG-3 antibody [CAL77] - BSA and Azide free

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAG-3 antibody [CAL77] - BSA and Azide free (AB251606)

Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling LAG-3 with ab237720 at 1/900 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining on the human tonsil is observed. The section was incubated with ab237720 for 15 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 a ready to use Goat Anti-Rabbit IgG H&L (HRP). Perform heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 30 mins.

This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237720).

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-LAG-3 antibody [CAL77] - BSA and Azide free (AB251606)

Intracellular flow cytometric analysis of 2% paraformaldehyde-fixed, 0.1%Tween-20 permeabilized human peripheral blood mononuclear cell (PBMC) treated with 10μg/ml PHA for 2 days, labeling Lymphocyte Activation Gene 3 with ab237720 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-CD3 conjugated to Alexa Fluor^® 647. Then fixed with 2% PFA for 10min followed by intracellular staining with rabbit IgG (Left) and ab237720 (Right).

This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237720).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAG-3 antibody [CAL77] - BSA and Azide free (AB251606)

Formalin-fixed, paraffin-embedded human tonsil tissue stained for LAG-3 with ab237720 at 0.25 μg/ml in immunohistochemical analysis.

This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237720).

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-LAG-3 antibody [CAL77] - BSA and Azide free (AB251606)

This data was developed using ab237720, the same antibody clone in a different buffer formulation.

Flow cytometry staining of human peripheral blood mononuclear cells (PBMCs) (top) or PBMCs treated with 5 μg/ml phytohaemagglutinin (PHA) for 48 hours (bottom), with ab237720 (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 on ice 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 ab237720 or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (1x 10⁶in 100 μl at 0.2 μg/ml (1/12750)) for 30 min at 22°C . The cells were simultaneously stained with CD3.

The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min at 22°C

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

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-LAG-3 antibody [CAL77] - BSA and Azide free (AB251606)

Intracellular flow cytometric analysis of 2% paraformaldehyde-fixed, 0.1%Tween-20 permeabilized mouse splenocytes treated with 2.5μg/ml Concanavalin A (ConA) for 3 days, labeling LAG-3 with ab237720 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-CD3 conjugated to Alexa Fluor^® 647. Then fixed with 2% PFA for 10min followed by intracellular staining with rabbit IgG (Left) and ab237720 (Right).

This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237720).