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 Mouse PD-1 Fc Chimera Protein, CF

Name: Recombinant Mouse PD-1 Fc Chimera Protein, CF
Brand: rnd
SKU: 1021-PD
Rating: 5 (1 reviews)

Catalog # 1021-PD

Citations (3)

(1)

Datasheet / COA / SDS

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$35 to United States

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Key Product Details

Source:	NS0
Accession #:	Q02242
Structure / Form:	Disulfide-linked homodimer
Applications:	Binding Activity

View Full Technical Details for Recombinant Mouse PD-1 Fc Chimera Protein, CF

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Product Specifications

Source

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

Mouse PD-1 (Leu25-Gln167) Accession # Q02242	IEGRMD	Human IgG₁ (Pro100-Lys330)
N-terminus		C-terminus

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.

N-terminal sequence Analysis

Leu25

Predicted Molecular Mass

43 kDa (monomer)

SDS-PAGE

66 kDa, reducing conditions

Activity

Measured by its binding ability in a functional ELISA. Immobilized Recombinant Mouse PD-1 Fc Chimera at 1 µg/mL (100 µL/well) can bind Recombinant Mouse B7‑H1/PD‑L1 Fc Chimera (Catalog # 1019-B7) with an ED₅₀ of 0.16-0.8 μg/mL.

Recombinant Mouse PD-1 Fc Chimera Protein, CF Scientific Data Examples

	Recombinant Mouse PD-1 Fc Chimera Protein Bioactivity When Recombinant Mouse PD-1 Fc Chimera (Catalog # 1021-PD) is coated at 1 µg/mL, Recombinant Mouse B7-H1/PD-L1 Fc Chimera (Catalog # 1019-B7) binds with an ED₅₀of 0.16-0.8 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.

1021-PD

Formulation	Lyophilized from a 0.2 μm filtered solution in Tris-HCL, NaCl and EDTA with Trehalose.
Reconstitution	Reconstitute at 0.5 mg/mL in sterile water. 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.

Volume (to add to vial)

Mass (in vial)

Desired Reconstitution Concentration

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 mouse PD-1 consists of a 149 amino acid (aa) extracellular region (ECD) with one immunoglobulin-like V-type domain, a 21 aa transmembrane domain, and a 98 aa cytoplasmic region. The mouse PD-1 ECD shares 65% aa sequence identity with the human 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).

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.
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 (3)

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Citations are publications that use Bio-Techne products. Selected citations for Recombinant Mouse PD-1 Fc Chimera Protein, CF include:

Species: Mouse
Sample Types: Protein
Applications: Enzyme Assay

Matsumura et al. (2015), Botulinum toxin A complex exploits intestinal M cells to enter the host and exert neurotoxicity. Nat Commun, 6(0):6255.
PMID: 25687350
Species: Mouse
Sample Types: Whole Cells
Applications: Flow Cytometry

Arnett HA et al. (2007), BTNL2, a butyrophilin/B7-like molecule, is a negative costimulatory molecule modulated in intestinal inflammation. J. Immunol., 178(3):1523-33.
PMID: 17237401
Species: Mouse
Sample Types: Whole Cells
Applications: Bioassay

Butte MJ et al. (2007), Programmed death-1 ligand 1 interacts specifically with the B7-1 costimulatory molecule to inhibit T cell responses. Immunity, 27(1):111-22.
PMID: 17629517

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Recombinant Mouse PD-1 Fc Chimera Protein, CF

Name: Anonymous

Application: Binding assay/Protein-protein interaction

Verified Customer | Posted 02/11/2019

0.5 ug mPD-1-hIgG1 (Cat# 1021-PD-100) was added to cells expressing mPD-L1 mCherry. Binding was detected with anti-human 488 secondary antibody.

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FAQs

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 Mouse PD-1 Fc Chimera Protein, CF

Order Information

Shipping Information

R&D Systems Guarantee

Key Product Details

Complete Your Experiment

Complete Your Research

Product Specifications

Recombinant Mouse PD-1 Fc Chimera Protein, CF Scientific Data Examples

Formulation, Preparation and Storage

Carrier Free

Reconstitution Calculator

Background: PD-1

Citations (3)

Customer Reviews (1)

Customer Images

Recombinant Mouse PD-1 Fc Chimera Protein, CF

FAQs

Product Datasheets

Product Datasheets

COA

Certificate of Analysis

SDS

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