The vial is supposed to contain lyophilized protein but it appears to be empty. Is there anything in it?

Pellets can be dislodged during shipping and become disbursed on the vial wall and in the cap. Centrifuge or tap the vial on the benchtop to return this material to the vial bottom. If this does not reveal a pellet, closely inspect the cone of the vial. Some pellets appear as only a tiny amount of material or as a transparent film due to the original buffer formulation. This is a normal appearance for many proteins. For example, if the product is originally lyophilized from a solvent such as acetonitrile or ethanol, and supplied carrier-free, you may not be able to detect the pellet with the naked eye. This does not mean the vial is empty. Reconstitute the vial as directed. After reconstitution, protein concentration can be tested with a spectrophotometer.

What is the recommended method for reconstitution of a lyophilized protein or antibody?

Unless more specific directions are on the Certificate of Analysis provided with the product, we suggest the following procedure to ensure optimal recovery: 1. Allow the vial and reconstitution buffer to equilibrate to room temperature. 2. Briefly centrifuge the vial to ensure that all lyophiliate is collected at the bottom of the vial. 3. Add the amount of buffer required to achieve the concentration recommended on the product insert. 4. Allow the vial to reconstitute for 15-30 minutes at room temperature with gentle agitation, like on a rocker platform or rotating by hand. Avoid vigorous shaking that can cause foaming and protein denaturation. 5. Aliquot into volumes greater than 20 μL and store as indicated on the product insert. If the vial exhibits flakes or particulates, mix the product for a couple of hours at room temperature and then at 4oC overnight. Contact Technical Service if product does not go into solution.

Are R&D Systems recombinant proteins and antibodies sterile?

Although the vials are bottled using aseptic techniques, heat-treated vials, and sterile stock solutions, they are not considered or guaranteed to be sterile. If sterile material is needed for an experiment, the material can be filtered through a 0.2 micron filter designed for use with biological fluids.

Recombinant Human PD-1 His Tagged Protein, CF

Name: Recombinant Human PD-1 His Tagged Protein, CF
Brand: R&D Systems
SKU: 8986-PD
Rating: 5 (10 reviews)

R&D Systems, Inc. a Bio-Techne Brand

Catalog # 8986-PD

Citations (7)

(10)

Product Datasheet / COA / SDS

Catalog #

Availability

Size / Price

Qty

Setting the standard in quality research reagents

Key Product Details

Source:	HEK293
Accession #:	Q15116.3
Applications:	Bioactivity

View Full Technical Details for Recombinant Human PD-1 His Tagged Protein, CF

Product Specifications for Recombinant Human PD-1 His Tagged Protein, CF

Source	Human embryonic kidney cell, HEK293-derived human PD-1 protein Leu25-Thr168, with a C-terminal 6-His tag
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.
N-terminal sequence Analysis	Leu25
Predicted Molecular Mass	17 kDa
SDS-PAGE	32 - 44 kDa, under reducing conditions.
Activity	Measured by its binding ability in a functional ELISA. When Recombinant Human PD-1 is immobilized at 1 µg/mL (100 µL/well), the concentration of Recombinant Human B7-H1/PD-L1 Fc Chimera (Catalog # 156-B7) that produces 50% of the optimal binding response is approximately 0.3-1.8 μg/mL.

Scientific Data Examples for Recombinant Human PD-1 His Tagged Protein, CF

	Bioactivity of Human PD-1 When recombinant human PD-1 (8986-PD) is immobilized at 1 µg/mL (100 µL/well), the concentration of recombinant human PD-L1 (B7-H1) Fc Chimera (156-B7) that produces 50% of the optimal binding response is approximately 0.3-1.8 μg/mL.

Formulation, Preparation and Storage

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.

8986-PD

Formulation	Lyophilized from a 0.2 μm filtered solution in PBS.
Reconstitution	Reconstitute at 400 μg/mL in PBS. Reconstitution Buffer Available: Size / Price Qty Loading... RB01:
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.

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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 148 amino acid (aa) extracellular region (ECD) with one immunoglobulin-like V-type domain, a 24 aa transmembrane domain, and a 95 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, 14).

Item	Value
References	Ishida, Y. et al. (1992) EMBO J. 11:3887. 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. Swaika, A. et al. (2015) Mol. Immunol. 67: 4
Long Name	Programmed Death-1
Entrez Gene IDs	5133 (Human); 18566 (Mouse); 301626 (Rat); 100533201 (Porcine); 486213 (Canine); 102123659 (Cynomolgus Monkey)
Alternate Names	CD279, CD279 antigen, PD-1, PD1, PD1hPD-l, PDCD1, Protein PD-1, SLEB2, hPD-1, programmed cell death 1, programmed cell death protein 1

Citations for Recombinant Human PD-1 His Tagged Protein, CF (7)

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Citations are publications that use Bio-Techne products. Selected citations for Recombinant Human PD-1 His Tagged Protein, CF include:

Species: Mouse
Sample Types: In Vivo
Applications: In Vivo

T Phakham et al. (2022), Highly efficient hybridoma generation and screening strategy for anti-PD-1 monoclonal antibody development Scientific Reports, 12(1):17792.
PMID: 36273231
Species: Human

Applications: Bioassay

CE Kang et al. (2022), Antibody-dependent cellular cytotoxicity-null effector developed using mammalian and plant GlycoDelete platform Scientific Reports, 12(1):19030.
PMID: 36347901
Species: Human
Sample Types: Protein
Applications: Bioassay

H Issafras et al. (2021), Structural basis of HLX10 PD-1 receptor recognition, a promising anti-PD-1 antibody clinical candidate for cancer immunotherapy PLoS ONE, 16(12):e0257972.
PMID: 34972111
Species: Human
Sample Types: Protein
Applications: Bioassay

CJ Edwards et al. (2021), The multi-specific VH-based Humabody CB213 co-targets PD1 and LAG3 on T cells to promote anti-tumour activity British Journal of Cancer, 0(0).
PMID: 34969998
Species: Human
Sample Types: Serum
Applications: Bioassay

M Metz et al. (2021), Fenebrutinib in H1 antihistamine-refractory chronic spontaneous urticaria: a randomized phase 2 trial Nature Medicine, 27(11):1961-1969.
PMID: 34750553
Species: Human
Sample Types: Protein
Applications: Surface Plasmon Resonance (SPR

M Crauwels et al. (2020), Reshaping Nanobodies for affinity purification on protein A N Biotechnol, 0(0).
PMID: 32001339
Species: Human
Sample Types: Reference Standard
Applications: Bioassay

ME Brown et al. (2020), Assessing the binding properties of the anti-PD-1 antibody landscape using label-free biosensors PLoS ONE, 15(3):e0229206.
PMID: 32134960

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Customer Reviews for Recombinant Human PD-1 His Tagged Protein, CF (10)

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Recombinant Human PD-1 His Tagged Protein, CF

Name: Anonymous

Application: SDS-PAGE Control

Verified Customer | Posted 10/15/2020
Recombinant Human PD-1 His Tagged Protein, CF

Name: Anonymous

Application: Binding assay/Protein-protein interaction

Reason for Rating: Good binding of PD-1-his tag with PDL1-Fc by Octet

Verified Customer | Posted 06/19/2020
Name: Jenna Nguyen

Application: Immunoassay Standard

Verified Customer | Posted 02/08/2019
Name: Jenna Nguyen

Application: SDS-PAGE Control

Verified Customer | Posted 01/30/2019
Recombinant Human PD-1 His Tagged Protein, CF

Name: Jenna Nguyen

Application: Cell migration/motility

Verified Customer | Posted 12/06/2018
Recombinant Human PD-1 His Tagged Protein, CF

Name: Jenna Nguyen

Application: CellProlif

Verified Customer | Posted 10/24/2018
Recombinant Human PD-1 His Tagged Protein, CF

Name: Jenna Nguyen

Application: Immunoassay Standard

Verified Customer | Posted 10/12/2018
Recombinant Human PD-1 His Tagged Protein, CF

Name: Jenna Nguyen

Application: Binding assay/Protein-protein interaction

Verified Customer | Posted 08/29/2018
Recombinant Human PD-1 His Tagged Protein, CF

Name: Casey Keyes

Application: Binding assay/Protein-protein interaction

Verified Customer | Posted 11/30/2017
Recombinant Human PD-1 His Tagged Protein, CF

Name: Anonymous

Application: Binding assay/Protein-protein interaction

Reason for Rating: This protein was used for a multitude of assays, including mouse immunizations, cell sorting, and Octet/binding assays (image of this assay included). The protein has performed well for all assays tested.

Verified Customer | Posted 05/17/2017

An anti-PD1 antibody was tested for binding to PD-1-His using ForteBio Octet, and the antibody bound specifically.

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FAQs for Recombinant Human PD-1 His Tagged Protein, CF

The vial is supposed to contain lyophilized protein but it appears to be empty. Is there anything in it?

Pellets can be dislodged during shipping and become disbursed on the vial wall and in the cap. Centrifuge or tap the vial on the benchtop to return this material to the vial bottom. If this does not reveal a pellet, closely inspect the cone of the vial. Some pellets appear as only a tiny amount of material or as a transparent film due to the original buffer formulation. This is a normal appearance for many proteins. For example, if the product is originally lyophilized from a solvent such as acetonitrile or ethanol, and supplied carrier-free, you may not be able to detect the pellet with the naked eye. This does not mean the vial is empty. Reconstitute the vial as directed. After reconstitution, protein concentration can be tested with a spectrophotometer.
What is the recommended method for reconstitution of a lyophilized protein or antibody?

Unless more specific directions are on the Certificate of Analysis provided with the product, we suggest the following procedure to ensure optimal recovery: 1. Allow the vial and reconstitution buffer to equilibrate to room temperature. 2. Briefly centrifuge the vial to ensure that all lyophiliate is collected at the bottom of the vial. 3. Add the amount of buffer required to achieve the concentration recommended on the product insert. 4. Allow the vial to reconstitute for 15-30 minutes at room temperature with gentle agitation, like on a rocker platform or rotating by hand. Avoid vigorous shaking that can cause foaming and protein denaturation. 5. Aliquot into volumes greater than 20 μL and store as indicated on the product insert. If the vial exhibits flakes or particulates, mix the product for a couple of hours at room temperature and then at 4^oC overnight. Contact Technical Service if product does not go into solution.
Are R&D Systems recombinant proteins and antibodies sterile?

Although the vials are bottled using aseptic techniques, heat-treated vials, and sterile stock solutions, they are not considered or guaranteed to be sterile. If sterile material is needed for an experiment, the material can be filtered through a 0.2 micron filter designed for use with biological fluids.

Recombinant Human PD-1 His Tagged Protein, CF

R&D Systems, Inc. a Bio-Techne Brand

Key Product Details

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Product Specifications for Recombinant Human PD-1 His Tagged Protein, CF

Scientific Data Examples for Recombinant Human PD-1 His Tagged Protein, CF

Formulation, Preparation and Storage

Carrier Free

Reconstitution Calculator

Background: PD-1

Citations for Recombinant Human PD-1 His Tagged Protein, CF (7)

Customer Reviews for Recombinant Human PD-1 His Tagged Protein, CF (10)

Customer Images

Recombinant Human PD-1 His Tagged Protein, CF

Recombinant Human PD-1 His Tagged Protein, CF

Recombinant Human PD-1 His Tagged Protein, CF

Recombinant Human PD-1 His Tagged Protein, CF

Recombinant Human PD-1 His Tagged Protein, CF

Recombinant Human PD-1 His Tagged Protein, CF

Recombinant Human PD-1 His Tagged Protein, CF

Recombinant Human PD-1 His Tagged Protein, CF

FAQs for Recombinant Human PD-1 His Tagged Protein, CF

Product Documents for Recombinant Human PD-1 His Tagged Protein, CF

Product Datasheets

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