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, mIHC |
| Clonality: | Monoclonal |
| Clone number: | PS01-33 |
| Form: | Liquid |
| Storage condition: | Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term. |
| 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 tonsils 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 mIHC |
1:200 1:100 1:1,000 1:400 |
| Uniprot #: | SwissProt: P18627 Human |
| Alternative names: | LAG3 FDC Lymphocyte activation gene 3 protein LAG-3 Protein FDC CD223 |
Images
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Fig1:
Immunohistochemical analysis of paraffin-embedded human tonsils 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. |
|
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. |
|
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. |
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Fig4: mIHC analysis of human tonsils tissue (Formalin/PFA-fixed paraffin-embedded sections) with Rabbit anti-LAG-3 antibody (HA721358) at 1/400 dilution. The immunostaining was performed with the IRISKit® HyperView mTSA Kit (MH900206). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 30 mins at 95℃. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Olympus VS200 Slide Scanner. |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.