Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
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Species reactivity: | Human, Mouse |
Applications: | WB, IHC-P, IF-Cell |
Clonality: | Monoclonal |
Clone number: | PSH0-44 |
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: | Recombinant protein within Human LAG-3 aa 1-500 / 525. |
Positive control: | NIH/3T3 transfected with LAG3 cell lysate, human tonsil tissue, NIH/3T3 transfected with LAG3. |
Subcellular location: | Cell membrane; Secreted. |
Recommended Dilutions:
WB IHC-P IF-Cell |
1:2,000 1:200 1:100 |
Uniprot #: | SwissProt: P18627 Human | Q61790 Mouse |
Alternative names: | CD223 CD223 antigen FDC protein LAG-3 Lag3 LAG3_HUMAN Lymphocyte activating 3 Lymphocyte activation gene 3 Lymphocyte activation gene 3 protein Protein FDC |
Fig1:
Western blot analysis of LAG-3 on different lysates with Rabbit anti-LAG-3 antibody (HA721346) at 1/2,000 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 kDa Observed band size: 55/70 kDa Exposure time: 1 minute 10 seconds; 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 (HA721346) at 1/2,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature. |
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Fig2:
Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-LAG-3 antibody (HA721346) 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 (HA721346) 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. |
Fig3:
Immunocytochemistry analysis of NIH/3T3 cells transfected with or without LAG3 labeling LAG-3 with Rabbit anti-LAG-3 antibody (HA721346) 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 (HA721346) 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. |