B7-H4 Antibody (PSH03-65)

Western Blot: B7-H4 Antibody (PSH03-65) [NBP3-32065]

Western Blot: B7-H4 Antibody (PSH03-65) [NBP3-32065] - Western blot analysis of B7-H4 on different lysates with Rabbit anti-B7-H4 antibody (NBP3-32065) at 1/1,000 dilution.

Lane 1: MDA-MB-468 cell lysate (no heat) (15 ug/Lane)
Lane 2: HeLa cell lysate (negative) (30 ug/Lane)
Lane 3: NIH:OVCAR-3 cell lysate (30 ug/Lane)
Lane 4: Mouse uterus tissue lysate (30 ug/Lane)
Lane 5: Rat ovary tissue lysate (15 ug/Lane)

Predicted band size: 31 kDa
Observed band size: 60-75 kDa

Exposure time: 35 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. The primary antibody (NBP3-32065) at 1/1,000 dilution was used in 5% NFDM/TBST at 4C overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.

Immunocytochemistry/ Immunofluorescence: B7-H4 Antibody (PSH03-65) [NBP3-32065]

Immunocytochemistry/ Immunofluorescence: B7-H4 Antibody (PSH03-65) [NBP3-32065] - Immunocytochemistry analysis of SK-Br-3 (positive) and HeLa (negative) labeling B7-H4 with Rabbit anti-B7-H4 antibody (NBP3-32065) at 1/1,000 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-H4 antibody (NBP3-32065) at 1/1,000 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.