Human PD-1 Quantikine ELISA Kit

Name: Human PD-1 Quantikine ELISA Kit
Brand: R&D Systems
SKU: DPD10
Price: 615 USD

Catalog # DPD10 | R&D Systems, Inc. a Bio-Techne Brand

Setting the standard in quality research reagents

Key Product Details

Assay Length:	4.5 hours
Sample Type & Volume Required Per Well:	Cell Culture Supernates (50 uL), Cell Lysates (50 uL), Serum (50 uL), EDTA Plasma (50 uL), Heparin Plasma (50 uL), Urine (50 uL)
Sensitivity:	3.27 pg/mL
Assay Range:	15.6 - 1,000 pg/mL (Cell Culture Supernates, Cell Lysates, Serum, EDTA Plasma, Heparin Plasma, Urine)
Human PD-1 Quantikine ELISA Kit Summary:	The Quantikine™ Human Programmed Death-1 (PD-1) Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human PD-1 in cell culture supernates, cell lysates, serum, plasma, and urine. It contains HEK293-expressed recombinant human PD-1 and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human PD-1 showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural human PD-1.

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Product Specifications for Human PD-1 Quantikine ELISA Kit

Assay Type	Solid Phase Sandwich ELISA
Format	96-well strip plate
Measurement	Quantitative ELISA
Detection Method	Colorimetric - 450nm (TMB)
Conjugate	HRP
Reactivity	Human, Cynomolgus Monkey
Specificity	Natural and recombinant human PD-1.
Cross-reactivity	< 0.5% cross-reactivity observed with available related molecules. < 50% cross-species reactivity observed with species tested.
Interference	No significant interference observed with available related molecules.

Precision

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision

Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in twenty separate assays to assess inter-assay precision.

Cell Culture Supernates, Cell Lysates, Serum, EDTA Plasma, Heparin Plasma, Urine

	Intra-Assay Precision			Inter-Assay Precision
Sample	1	2	3	1	2	3
n	20	20	20	20	20	20
Mean (pg/mL)	125	319	655	131	304	626
Standard Deviation	3.49	5.6	10.8	7.36	14.6	25.8
CV%	2.6	1.8	1.6	5.6	4.8	4.1

Recovery

The recovery of human PD-1 spiked to levels throughout the range of the assay was evaluated.

Sample Type	Average % Recovery	Range %
Cell Culture Media (n=4)	100	97-103
Cell Lysis Buffer (n=1)	96	94-97
EDTA Plasma (n=4)	86	81-93
Heparin Plasma (n=4)	85	75-93
Serum (n=4)	88	77-98
Urine (n=4)	89	83-97

Linearity

To assess the linearity of the assay, samples containing and/or spiked with high concentrations of human PD-1 were serially diluted with calibrator diluent to produce samples with values within the dynamic range of the assay.

Scientific Data Examples for Human PD-1 Quantikine ELISA Kit

	Human PD-1 ELISA Standard Curve
	Human PD-1 Quantikine Recovery Competitor Comparison PD-1 is spiked at three known concentrations throughout the range of the assay and run to measure response of the spiked sample matrix. Serum recovery is 95% compared to 136% for the top competitor. EDTA Plasma recovery is 87% compared to 157% for the top competitor. Culture media recovery is 94% compared to 58% for the top competitor. In spike and recovery experiments, natural samples are spiked with the recombinant target analyte of interest to identify interference caused by sample matrices.

Preparation and Storage

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 2 - 8 °C. Do not use past expiration date.

Background: PD-1

Programmed Death-1 (PD-1), also known as Programmed cell death protein and CD279, is an extensively studied immune checkpoint inhibitory receptor. Given PD-1's role in peripheral tolerance, it is not surprising that increased PD-1 expression is a mechanism for immune escape, which is permissive for cancer growth and metastasis (1,2). PD‐1 is encoded by the PDCD1 gene (3). The PD-1 glycoprotein is a monomeric 50-55kDa type 1 transmembrane protein that belongs to the immunoglobulin (Ig) superfamily (4). PD-1 expression and induction have been well studied (3,5,6). It is expressed in CD4⁺ and CD8⁺ T cells as well as B cells, macrophages, some dendritic cell subsets and NK cells. PD-1 is induced by T cell receptor (TCR) signaling as well as interleukin 2 (IL-2), IL-7 and type 1 interferons. A variety of reagents are used to experimentally induce PD-1 expression, including, phorbol 12-myristate 13 acetate, ionomycin, concanavalin A, CD3/CD28 antibodies, or most notably lymphocytic choriomeningitis virus in vivo (6).

PD-1 expression is also regulated post-translationally (2). The PD-1 extracellular domain is glycosylated at asparagine (N) residues N49 and N72 while in the endoplasmic reticulum. PD-1 subsequently transits to the golgi apparatus where it is fucosylated at the same sites by the core fucosylase FUT8. This post translational modification represents a novel therapeutic target as T cells have a more robust anticancer response due to reduced surface expression of de-fucosylated PD-1. PD-1 is polyubiquinated at lysine (K) residue K48 by the E3 ligase F-Box 38 (FBX028) (2,7) which results in PD-1 degradation via the proteosome.

PD-1 serves as the receptor for programmed death ligand 1 (PD-L1/CD274/B7-H1) and PD-L2 (CD273/B7-DC) (8,9). The interaction of the extracellular domains of PD-1 and PD-L1 causes a confirmational change that results in the phosphorylation of the cytoplasmic immunoreceptor tyrosine-based inhibitory motif (ITIM) and the immunoreceptor tyrosine-based switch motif (ITSM) by Src family kinases (4). Src homology 2 containing tyrosine phosphatase 2 (SHP-2) and SHP-1 are recruited in order to attenuate T cell activating signals via dephosphorylation of downstream signaling cascades.

Item	Value
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

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Human PD-1 Quantikine ELISA Kit

Catalog # DPD10 | R&D Systems, Inc. a Bio-Techne Brand

Key Product Details

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Product Specifications for Human PD-1 Quantikine ELISA Kit

Precision

Recovery

Linearity

Scientific Data Examples for Human PD-1 Quantikine ELISA Kit

Preparation and Storage

Background: PD-1

Citations for Human PD-1 Quantikine ELISA Kit

Product Documents for Human PD-1 Quantikine ELISA Kit

Product Datasheets

COA

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