LAG-3 Recombinant Rabbit Monoclonal Antibody [PS01-33]
cat.: HA721358
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human
Applications: IHC-P, IF-Cell, WB
Clonality: Monoclonal
Clone number: PS01-33
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 57 kDa
Isotype: IgG
Immunogen: Synthetic peptide.
Positive control: Human tonsil tissue, NIH/3T3 transfected with LAG3 cell lysate, NIH/3T3 cells transfected with LAG-3.
Subcellular location: Cell membrane; Secreted.
Recommended Dilutions:
  IHC-P
  IF-Cell
  WB

1:200
1:100
1:1,000
Uniprot #: SwissProt: P18627 Human
Alternative names: LAG3 FDC Lymphocyte activation gene 3 protein LAG-3 Protein FDC CD223
Images
HA721358_1.jpg Fig1: Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-LAG-3 antibody (HA721358) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721358) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA721358_2.jpg Fig2: Western blot analysis of LAG-3 on different lysates with Rabbit anti-LAG-3 antibody (HA721358) at 1/1,000 dilution.

Lane 1: NIH/3T3 transfected with LAG3 expression vector, whole cell lysate
Lane 2: NIH/3T3 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 57.4 kDa
Observed band size: 57/70 kDa

Exposure time: 2 minutes;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721358) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.
HA721358_3.jpg Fig3: Immunocytochemistry analysis of NIH/3T3 cells transfected with or without LAG-3 labeling LAG-3 with Rabbit anti-LAG-3 antibody (HA721358) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes, and then blocked with 1% BSA for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-LAG-3 antibody (HA721358) at 1/100 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.