Recombinant Human PD-L1/B7-H1 Fc Chimera Biotin Protein, CF

Catalog #: BT156 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
BT156-050

Recombinant Human PD-L1/B7-H1 Fc Chimera Biotin Protein Binding Activity

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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Human PD-L1/B7-H1 Fc Chimera Biotin Protein, CF Summary

Product Specifications

Purity

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<0.10 EU per 1 μg of the protein by the LAL method.

Activity

Measured by its binding ability in a functional ELISA. When Recombinant Human PD‑1 Fc Chimera (Catalog # 1086-PD) is immobilized at 1 μg/mL, 100 μL/well, the concentration of Biotinylated Recombinant PD-L1/B7-H1 Fc Chimera (Catalog# BT156) that produces 50% of the optimal binding response is 10-60 ng/mL.

Source

Mouse myeloma cell line, NS0-derived human PD-L1/B7-H1 protein

Human PD-L1 (Phe19-Thr239) Accession # Q9NZQ7	DIEGRMD	Human IgG₁ (Pro100-Lys330)
N-terminus		C-terminus

Accession #

Q9NZQ7

N-terminal Sequence
Analysis

Phe19

Structure / Form

Disulfide-linked homodimer, biotinylated via amines.

Predicted Molecular Mass

52 kDa

SDS-PAGE

70-75 kDa, reducing conditions

Product Datasheets

BT156

Product Datasheet

COA

SDS

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

BT156

Formulation	Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
Reconstitution	Reconstitute at 500 μg/mL in PBS.
Shipping	The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
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. 3 months, -20 to -70 °C under sterile conditions after reconstitution.

Scientific Data

Binding Activity

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When Recombinant Human PD-1 Fc Chimera (Catalog # 1086-PD) is immobilized at 1 µg/mL (100 µL/well), Biotinylated Recombinant Human PD-L1/B7-H1 Fc Chimera (Catalog # BT156) binds with an ED₅₀ of 10-60 ng/mL.

SDS-PAGE

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2 μg/lane of Biotinylated Recombinant Human PD-L1/B7-H1 Fc Chimera (Catalog # BT156) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 70-75 kDa and 140-150 kDa, respectively.

Reconstitution Calculator

Background: PD-L1/B7-H1

B7-H1, also known as PD-L1 and CD274, is an approximately 65 kDa transmembrane glycoprotein in the B7 family of immune regulatory molecules (1). Mature human B7-H1 consists of a 220 amino acid (aa) extracellular domain (ECD) with two immunoglobulin-like domains, a 21 aa transmembrane segment, and a 31 aa cytoplasmic domain (2). Within the ECD, human B7-H1 shares 73% and 74% aa sequence identity with mouse and rat B7-H1, respectively. Alternative splicing generates additional isoforms that either lack the first Ig-like domain or are truncated within the second Ig-like domain (3). B7-H1 is expressed on
inflammatory-activated immune cells including macrophages, T cells, and B cells (4-7), keratinocytes (8, 9), enothelial and intestinal epithelial cells (8, 10), as well as a variety of carcinomas and melanoma (11, 12). B7-H1 binds to T cell B7-1/CD80 and PD-1 (7, 8, 12-15). It suppresses T cell activation and proliferation (5, 8, 14, 16) and induces the apoptosis of activated T cells (11). It plays a role in the development of immune tolerance by promoting T cell anergy (7, 14) and enhancing regulatory T cell development (16). B7-H1 favors the development of anti-inflammatory IL-10 and IL-22 producing dendritic cells (5, 10) and inhibits the development of Th17 cells (16). In cancer, B7-H1 provides resistance to T cell mediated lysis, enhances EMT, and enhances the tumorigenic function of Th22 cells (6, 9, 12, 15).

References

Ceeraz, S. et al. (2013) Trends Immunol. 34:556.
Dong, H. et al. (1999) Nat. Med. 5:1365.
Frigola, X. et al. (2011) Clin. Cancer Res. 17:1915.
Tamura, H. et al. (2001) Blood 97:1809.
Chen, L. et al. (2007) J. Immunol. 178:6634.
Kuang, D.-M. et al. (2014) J. Clin. Invest. 124:4657.
Tsushima, F. et al. (2007) Blood 110:180.
Mazanet, M.M. and C.C.W. Hughes (2002) J. Immunol. 169:3581.
Cao, Y. et al. (2010) Cancer Res. 71:1235.
Scandiuzzi, L. et al. (2014) Cell Rep. 6:625.
Dong, H. et al. (2002) Nat. Med. 8:793.
Azuma, T. et al. (2008) Blood 111:3635.
Butte, M.J. et al. (2008) Mol. Immunol. 45:3567.
Park, J.-J. et al. (2010) Blood 116:1291.
Ritprajak, P. et al. (2010) J. Immunol. 184:4918.
Herold, M. et al. (2015) J. Immunol. 195:3584.

Long Name

Programmed Death Ligand 1

Entrez Gene IDs

29126 (Human); 60533 (Mouse); 499342 (Rat); 574058 (Porcine); 484186 (Canine); 102145573 (Cynomolgus Monkey)

Alternate Names

Avelumab; B7-H; B7H1; B7-H1; B7H1PDCD1L1; CD274 antigenMGC142294; CD274 molecule; CD274; PDCD1L1; PDCD1LG1; PDCD1LG1MGC142296; PDL1; PD-L1; PD-L1B7 homolog 1; PDL1PDCD1 ligand 1; programmed cell death 1 ligand 1; Programmed death ligand 1