B7-H3/CD276 Antibody (PSH02-46)

Immunocytochemistry/ Immunofluorescence: B7-H3/CD276 Antibody (PSH02-46) [NBP3-32139]

Immunocytochemistry/ Immunofluorescence: B7-H3/CD276 Antibody (PSH02-46) [NBP3-32139] - Immunocytochemistry analysis of THP-1 cells labeling B7-H3/CD276 with Rabbit anti-B7-H3/CD276 antibody (NBP3-32139) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-B7-H3/CD276 antibody (NBP3-32139) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 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 (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.

Flow Cytometry: B7-H3/CD276 Antibody (PSH02-46) [NBP3-32139]

Flow Cytometry: B7-H3/CD276 Antibody (PSH02-46) [NBP3-32139] - Flow cytometric analysis of THP-1 cells labeling B7-H3/CD276.

Cells were washed twice with cold PBS and resuspend. Then stained with the primary antibody (NBP3-32139, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody 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: B7-H3/CD276 Antibody (PSH02-46) [NBP3-32139]

Western blot analysis on different lysates with NBP3-32139 at 1/2,000 dilution. Lane 1: HeLa cell lysate Lane 2: MCF7 cell lysate Lane 3: HEK-293 cell lysate Lane 4: LoVo cell lysate Lane 5: U-2 OS cell lysate Lane 6: LNCaP cell lysate Lane 7: SH-SY5Y cell lysate Lane 8: THP-1 cell lysate Lane 9: HCT 116 cell lysate Lane 10: Raji cell lysate (negative) Lysates/proteins at 20 µg/Lane. Predicted band size: 57 kDa Observed band size: 100 kDa Exposure time: 42 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. NBP3-32139 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.