WB
Western blot analysis of SLP76 on different lysates with Rabbit anti-SLP76 antibody at 1/2,000 dilution. Lane 1: THP-1 cell lysate, Lane 2: HeLa cell lysate (negative), Lane 3: HL-60 cell lysate, Lysates/proteins at 20 µg/Lane. Exposure time: 3 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 at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-SLP76 antibody at 1/5,000 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 1/5,000 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 THP-1 (positive) and HeLa (negative) labeling SLP76 with Rabbit anti-SLP76 antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-SLP76 antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. 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.IP
SLP76 was immunoprecipitated from 0.2 mg HL-60 cell lysate with Rabbit anti-SLP76 antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using Rabbit anti-SLP76 antibody at 1/1,000 dilution. Mouse Anti-Rabbit IgG kappa light chain secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: HL-60 cell lysate (input) Lane 2: Rabbit anti-SLP76 antibody IP in HL-60 cell lysate Lane 3: Rabbit IgG instead of Rabbit anti-SLP76 antibody in HL-60 cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 14 seconds.| Product Name | SLP76 Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Recombinant protein within human SLP76 aa 1- 533/533 |
| Clonality | monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | ICC/IFIHCIPWB |
| WB:1:2000 IHC:1:5000 ICC/IF:1:100 IP:1-2μg/sample |
|
| Species Reactivity | Human |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | LCP2 |
|---|---|
| Gene Synonyms | IMD81 SLP76 SLP-76 |
| Gene Full Name | lymphocyte cytosolic protein 2 |
| Gene Summary | This gene encodes an adapter protein that acts as a substrate of the T cell antigen receptor (TCR)-activated protein tyrosine kinase pathway. The encoded protein associates with growth factor receptor bound protein 2, and is thought to play a role TCR-mediated intracellular signal transduction. A similar protein in mouse plays a role in normal T-cell development and activation. Mice lacking this gene show subcutaneous and intraperitoneal fetal hemorrhaging, dysfunctional platelets and impaired viability. [provided by RefSeq, Nov 2016] |
| Molecular Weight(MW) | 60kDa（Observed band size: 75kDa） |
| Cellular Localization | Cytoplasm. |
WB
Western blot analysis of SLP76 on different lysates with Rabbit anti-SLP76 antibody at 1/2,000 dilution. Lane 1: THP-1 cell lysate, Lane 2: HeLa cell lysate (negative), Lane 3: HL-60 cell lysate, Lysates/proteins at 20 µg/Lane. Exposure time: 3 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 at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-SLP76 antibody at 1/5,000 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 1/5,000 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 THP-1 (positive) and HeLa (negative) labeling SLP76 with Rabbit anti-SLP76 antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-SLP76 antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. 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.
IP
SLP76 was immunoprecipitated from 0.2 mg HL-60 cell lysate with Rabbit anti-SLP76 antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using Rabbit anti-SLP76 antibody at 1/1,000 dilution. Mouse Anti-Rabbit IgG kappa light chain secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: HL-60 cell lysate (input) Lane 2: Rabbit anti-SLP76 antibody IP in HL-60 cell lysate Lane 3: Rabbit IgG instead of Rabbit anti-SLP76 antibody in HL-60 cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 14 seconds.| Application Notes | WB:1:2000 IHC:1:5000 ICC/IF:1:100 IP:1-2μg/sample |
|---|
| 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 OM644057