WB
Western blot analysis of LAG-3 on different lysates with Rabbit anti-LAG-3 antibody 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. 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 at 1/2,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:200,000 dilution was used for 1 hour at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-LAG-3 antibody 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 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.ICC/IF
Immunocytochemistry analysis of NIH/3T3 cells transfected with or without LAG3 labeling LAG-3 with Rabbit anti-LAG-3 antibody 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 at 1/100 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.| Product Name | LAG-3 Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Recombinant protein within Human LAG-3 aa 1-500 / 525. |
| Clonality | monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | ICC/IFIHCWB |
| WB:1:2000 IHC:1:200 ICC/IF:1:100 |
|
| Species Reactivity | HumanMouse |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | LAG3 |
|---|---|
| Gene Synonyms | CD223 |
| Gene Full Name | lymphocyte activating 3 |
| Gene Summary | Lymphocyte-activation protein 3 belongs to Ig superfamily and contains 4 extracellular Ig-like domains. The LAG3 gene contains 8 exons. The sequence data, exon/intron organization, and chromosomal localization all indicate a close relationship of LAG3 to CD4. [provided by RefSeq, Jul 2008] |
| Molecular Weight(MW) | 57kDa(Observed band size:55/70kDa) |
| Cellular Localization | Cell membrane; Secreted. |
WB
Western blot analysis of LAG-3 on different lysates with Rabbit anti-LAG-3 antibody 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. 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 at 1/2,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:200,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-LAG-3 antibody 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 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.
ICC/IF
Immunocytochemistry analysis of NIH/3T3 cells transfected with or without LAG3 labeling LAG-3 with Rabbit anti-LAG-3 antibody 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 at 1/100 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.| Application Notes | WB:1:2000 IHC:1:200 ICC/IF:1:100 |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
Data sheet for OM644041