Recombinant Human PD-1 Fc Chimera Biotinylated Protein, CF Summary
Product Specifications
Analysis
Product Datasheets
Carrier Free
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.
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: | Store the unopened product at -20 to -70 °C. Use a manual defrost freezer and avoid repeated freeze-thaw cycles. Do not use past expiration date. |
Scientific Data
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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.
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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).
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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).
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- Sharpe, A.H. and G. J. Freeman (2002) Nat. Rev. Immunol. 2:116.
- Coyle, A. and J. Gutierrez-Ramos (2001) Nat. Immunol. 2:203.
- Nishimura, H. and T. Honjo (2001) Trends Immunol. 22:265.
- Watanabe, N et al. (2003) Nat. Immunol. 4:670.
- Zhang, X. et al. (2004) Immunity 20:337.
- Lázár-Molnár, E. et al. (2008) Proc. Natl. Acad. Sci. USA 105:10483.
- Nishimura, H et al. (1996) Int. Immunol. 8:773.
- Keir, M.E. et al. (2008) Annu. Rev. Immunol. 26:677.
- Butte, M.J. et al. (2007) Immunity 27:111.
- Okazaki, T. et al. (2013) Nat. Immunol. 14:1212.
- Iwai, Y. et al. (2002) Proc. Natl. Acad. Sci. USA 99: 12293.
- Nogrady, B. (2014) Nature 513:S10.
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