Recombinant Human PD-1 Fc Chimera Biotinylated Protein, CF

Catalog #: BT1086 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
BT1086-050

Recombinant Human PD-1 Fc Chimera Biotinylated Protein Binding Activity

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

Recombinant Human PD-1 Fc Chimera Biotinylated Protein, CF Summary

Product Specifications

Purity

>90%, 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 B7-H1/PD-L1 Fc Chimera (Catalog # 156-B7) is immobilized at 0.5 µg/mL (100 µL/well), Biotinylated Recombinant Human PD-1 Fc Chimera (Catalog # BT1086) binds with an ED₅₀ of 10.0-100 ng/mL.

Source

Mouse myeloma cell line, NS0-derived human PD-1 protein

Human PD-1 (Leu25-Gln167) Accession # Q15116.3	IEGRMD	Human IgG₁ (Pro100-Lys330)
N-terminus		C-terminus

Accession #

Q15116.3

N-terminal Sequence
Analysis

Leu25

Structure / Form

Disulfide-linked homodimer, biotinylated proteins via amines.

Predicted Molecular Mass

43 kDa

SDS-PAGE

60-70 kDa, under reducing conditions.

Product Datasheets

BT1086

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.

BT1086

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 B7-H1/PD-L1 Fc Chimera (156-B7) is coated at 0.5 µg/mL, 100 µL/well, Biotinylated Recombinant Human PD-1 Fc Chimera (Catalog # BT1086) binds with an ED₅₀ of 10.0-100 ng/mL.

Flow Cytometry

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In a functional flow cytometry test, (A) Recombinant Human PD-1 Fc Chimera Biotinylated Protein (Catalog # BT1086) binds to HEK293 human embryonic kidney cell line transfected with recombinant human PD-L1 and EGFP. Ligand binding was detected by staining cells with APC-conjugated Streptavidin (F0050), which does not stain the cells in the absence of recombinant protein (B).

SDS-PAGE

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2 μg/lane of Recombinant Biotinylated Human PD‑1 Fc Chimera was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 60-70 kDa and 120-140 kDa, respectively.

Reconstitution Calculator

Background: PD-1

Programmed Death-1 receptor (PD-1), also known as CD279, is type I transmembrane protein belonging to the CD28 family of immune regulatory receptors (1). Other members of this family include CD28, CTLA-4, ICOS, and BTLA (2-5). Mature human PD-1 consists of a 150 amino acid (aa) extracellular region (ECD) with one immunoglobulin-like V-type domain, a 21 aa transmembrane domain, and a 97 aa cytoplasmic region. The human PD-1 ECD shares 65% aa sequence identity with the mouse PD-1 ECD. The cytoplasmic tail contains two tyrosine residues that form the immunoreceptor tyrosine-based inhibitory motif (ITIM) and immunoreceptor tyrosine-based switch motif (ITSM) that are important for mediating PD-1 signaling. PD-1 acts as a monomeric receptor and interacts in a 1:1 stoichiometric ratio with its ligands PD-L1 (B7-H1) and PD-L2 (B7-DC) (6, 7). PD‑1 is expressed on activated T cells, B cells, monocytes, and dendritic cells while PD-L1 expression is constitutive on the same cells and also on nonhematopoietic cells such as lung endothelial cells and hepatocytes (8, 9). Ligation of PD-L1 with PD-1 induces co‑inhibitory signals on T cells promoting their apoptosis, anergy, and functional exhaustion (10). Thus, the PD-1:PD-L1 interaction is a key regulator of the threshold of immune response and peripheral immune tolerance (11). Finally, blockade of the PD-1: PD-L1 interaction by either antibodies or genetic manipulation accelerates tumor eradication and shows potential for improving cancer immunotherapy (12, 13).

References