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Western blot analysis of CD161 on different lysates with Mouse anti-CD161 antibody (HA601063) at 1/2,000 dilution.
Lane 1: THP-1 cell lysate (20 µg/Lane)
Lane 2: HL-60 cell lysate (20 µg/Lane)
Lane 3: K-562 cell lysate (20 µg/Lane)
Lane 4: Jurkat cell lysate (20 µg/Lane)
Lane 5: Human kidney tissue lysate (40 µg/Lane)
Predicted band size: 25 kDa
Observed band size: 25 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 (HA601063) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Anti-Mouse IgG for IP Nano-secondary antibody (NBI02H) at 1/5,000 dilution was used for 1 hour at room temperature.
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Immunohistochemical analysis of paraffin-embedded human kidney tissue with Mouse anti-CD161 antibody (HA601063) at 1/500 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 (HA601063) at 1/500 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.
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Flow cytometric analysis of THP-1 cells labeling CD161.
Cells were washed twice with cold PBS and resuspend. Then stained with the primary antibody (HA601063, 1ug/ml) (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).
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