SIRP alpha Mouse Monoclonal Antibody [A3E3]

Immunocytochemistry analysis of THP-1 cells labeling SIRP alpha with Mouse anti-SIRP alpha antibody (EM1902-37) at 1/50 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 Mouse anti-SIRP alpha antibody (EM1902-37) at 1/50 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) 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 (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution.

Immunohistochemical analysis of paraffin-embedded human lung cancer tissue using anti-SIRP alpha antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (EM1902-37, 1/100) for 30 minutes 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.

Flow cytometric analysis of THP-1 cells labeling SIRP alpha.

Cells were washed twice with cold PBS and resuspend. Then stained with the primary antibody (EM1902-37, 1/1,000) (red) compared with Mouse IgG1 Isotype Control (green). After incubation of the primary antibody at +4℃ for 30 minutes, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Mouse IgG Secondary antibody (HA1125) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

Western blot analysis of SIRP alpha on different lysates with Mouse anti-SIRP alpha antibody (EM1902-37) at 1/1,000 dilution.

Lane 1: U-937 (Human acute monocytic leukemia cells) cell lysate
Lane 2: THP-1 (Human lymphoid lymphoma cells) cell lysate

Exposure time: 42 seconds

Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: EM1902-37, 1/1,000 in 5% NFDM/TBST, overnight at 4 ℃
Secondary antibody: Goat anti-Mouse IgG-HRP (HA1006), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

Predicted band size: 55.0 kDa
Observed band size: 80 kDa