Detection of Mouse B7‑H1/PD‑L1 by Western Blot. Western blot shows lysates of RAW 264.7 mouse monocyte/macrophage cell line untreated (-) or treated (+) with |
10 μg/mL LPS for 4 hours. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Mouse B7‑H1/PD‑L1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1019) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for B7‑H1/PD‑L1 at approximately 50-55 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
|Detection of B7‑H1/PD‑L1 in RAW 264.7 Mouse Cell Line by Flow Cytometry. RAW 264.7 mouse monocyte/macrophage cell line either treated with LPS overnight (filled histogram) or untreated (open histogram) was stained with Goat Anti-Mouse B7‑H1/PD‑L1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1019), followed by Allophycocyanin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0108). View our protocol for Staining Membrane-associated Proteins.|
|Detection of B7‑H1/PD‑L1 in HEK293 Human Cell Line Transfected with Mouse B7-H1/PD-L1 and eGFP by Flow Cytometry. HEK293 human embryonic kidney cell line transfected with either (A) mouse B7-H1/PD-L1 or (B) irrelevant transfectants and eGFP was stained with Goat Anti-Mouse B7‑H1/PD‑L1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1019) followed by Allophycocyanin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0108). Quadrant markers were set based on control antibody staining (Catalog # AB-108-C). View our protocol for Staining Membrane-associated Proteins.|
Mouse B7 homolog 1(B7-H1), also called programmed death ligand 1 (PD-L1) and programmed cell death 1 ligand 1 (PDCD1L1), is a member of the B7 family of proteins that provide signals for regulating T-cell activation and tolerance (1‑4). Other family members include B7-1, B7-2, B7-H2, B7-H3 and PD-L2. B7 proteins are immunoglobulin (Ig) superfamily members with extracellular Ig-V-like and Ig-C-like domains and a short cytoplasmic region. Among the family members, they share from 20‑40% amino acid (aa) sequence identity. The cloned mouse B7-H1/PD-L1 cDNA encodes a 290 aa type I membrane precursor protein with a putative 18 aa signal peptide, a 220 aa extracellular region containing one V-like and one C-like Ig domain, a 22 aa transmembrane region, and a 30 aa cytoplasmic domain. Mouse and human B7-H1/PD-L1 share approximately 70% aa sequence identity. B7-H1/PD-L1 is one of two ligands for programmed death-1 (PD-1), a member of the CD28 family of immunoreceptors. The other identified ligand is PD-L2. Mouse B7-H1/PD-L1 and PD-L2 share approximately 34% aa sequence identity and have similar functions. B7-H1/PD-L1 is constitutively expressed in various lymphoid and non-lymphoid organs including placenta, heart, pancreas, lung, liver, and endothelium (1‑4). The expression of B7-H1/PD-L1 is detected on B cells, T cells, monocytes, dendritic cells and thymic epithelial cells. IFN-gamma treatment induces B7-H1/PD-L1 expression in monocytes, dendritic cells, and endothelial cells. B7-H1/PD-L1 expression is also upregulated in a variety of tumor cell lines. On previously activated T cells, B7-H1/PD-L1 interaction with PD-1 inhibits TCR-mediated proliferation and cytokine production, suggesting an inhibitory role in regulating immune responses. In contrast, a costimulatory function for the PD-1 ligands on resting T cells has also been reported (1‑4).
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