CD315 Mouse Monoclonal Antibody [4E9G7]

Western blot analysis of CD315 against human CD315 (AA: extra 46-221) recombinant protein. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (EM1712-27, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody at 1:5,000 dilution was used for 1 hour at room temperature.

Western blot analysis of CD315 against HEK293 (1) and CD315 (AA: extra 46-221)-hIgGFc transfected HEK293 (2) cell lysate. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (EM1712-27, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody at 1:5,000 dilution was used for 1 hour at room temperature.

Immunohistochemical analysis of paraffin-embedded cervical cancer tissues using anti-CD315 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0) 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 (EM1712-27, 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 CD315 was done on HL-60 cells. The cells were fixed, permeabilized and stained with the primary antibody (EM1712-27, 1/100) (green). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated goat anti-Mouse IgG Secondary antibody at 1/500 dilution for 30 minutes. Unlabelled sample was used as a control (cells without incubation with primary antibody; red).