Detection of PD-L1/B7-H1 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 historgram) was stained with Rat Anti-Mouse PD-L1/B7-H1 Monoclonal Antibody (Catalog # MAB9078), followed by Allophycocyanin-conjugated Anti-Rat IgG Secondary Antibody (Catalog # F0113). View our protocol for Staining Membrane-associated Proteins.
Detection ofPD-L1/B7-H1 in HEK293 Human Cell Line Transfected with Mouse PD-L1/B7-H1 and eGFP by Flow Cytometry.
HEK293 human embryonic kidney cell line transfected with either (A) mouse PD-L1/B7-H1 or (B) irrelevant transfectants and eGFP was stained with and either (A) Rat Anti-Mouse PD-L1/B7-H1 Monoclonal Antibody (Catalog # MAB9078) or (B) Rat IgG2A Isotype Control (Catalog # MAB006) followed by Phycoerythrin-conjugated Anti-Hamster IgG Secondary Antibody .
Preparation and Storage
Reconstitute at 0.5 mg/mL in sterile PBS.
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
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).
Tamura, H. et al. (2001) Blood 97:1809.
Freeman, G. et al. (2000) J. Exp. Med. 192:1027.
Sharpe, A.H. and G. J. Freeman (2002) Nat. Rev. Immunol. 2:116.
Coyle, A. and J. Gutierrez-Ramos (2001) Nat. Immunol. 2:203.
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