SLP76 Recombinant Rabbit Monoclonal Antibody [PSH09-22]

☑ Relative expression (RE)

Western blot analysis of SLP76 on different lysates with Rabbit anti-SLP76 antibody (HA723080) 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.

Predicted band size: 60 kDa
Observed band size: 75 kDa

Exposure time: 3 minutes; ECL: K1801;

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 (HA723080) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

☑ Relative expression (RE)

Immunocytochemistry analysis of THP-1 (positive) and HeLa (negative) labeling SLP76 with Rabbit anti-SLP76 antibody (HA723080) 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 (HA723080) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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 (HA601187, 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.

Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-SLP76 antibody (HA723080) 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 (HA723080) 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.

Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-SLP76 antibody (HA723080) 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 (HA723080) 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.

SLP76 was immunoprecipitated from 0.2 mg HL-60 cell lysate with HA723080 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA723080 at 1/1,000 dilution. Mouse Anti-Rabbit IgG kappa light chain secondary antibody (M1208-2) at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: HL-60 cell lysate (input)
Lane 2: HA723080 IP in HL-60 cell lysate
Lane 3: Rabbit IgG instead of HA723080 in HL-60 cell lysate

Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 14 seconds; ECL: K1801