Negatively regulates T-cell-mediated immune response by inhibiting T-cell activation, proliferation, cytokine production and development of cytotoxicity. When expressed on the cell surface of tumor macrophages, plays an important role, together with regulatory T-cells (Treg), in the suppression of tumor-associated antigen-specific T-cell immunity. Involved in promoting epithelial cell transformation.
Background References
1. Chen L et al. The B7H4-PDL1 classifier stratifies immuno-phenotype in cervical cancer. Cancer Cell Int. 2022 Jan
2. Peuker K et al. Microbiota-dependent activation of the myeloid calcineurin-NFAT pathway inhibits B7H3- and B7H4-dependent anti-tumor immunity in colorectal cancer. Immunity. 2022 Apr
Western blot analysis of B7H4 on different lysates with Rabbit anti-B7H4 antibody (HA722024) at 1/1,000 dilution.
Lane 1: MDA-MB-468 cell lysate (no heat) (15 µg/Lane) Lane 2: HeLa cell lysate (negative) (30 µg/Lane) Lane 3: NIH:OVCAR-3 cell lysate (30 µg/Lane) Lane 4: Mouse uterus tissue lysate (30 µg/Lane) Lane 5: Rat ovary tissue lysate (15 µg/Lane)
Notice: no heat means the lysate is not boiled.
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 (HA722024) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
☑ Relative expression (RE)
Immunocytochemistry analysis of SK-Br-3 (positive) and HeLa (negative) labeling B7H4 with Rabbit anti-B7H4 antibody (HA722024) 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-B7H4 antibody (HA722024) at 1/1,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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 (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
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