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Human PD-1 QuicKit ELISA

Bio-Techne includes R&D Systems | Catalog # QK1086

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QK1086

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Key Product Details

Assay Length

80 minutes

Sample Type & Volume Required Per Well

Cell Culture Supernates (50 µL), Cell Lysates (50 µL), Serum (50 µL), EDTA Plasma (50 µL), Heparin Plasma (50 µL)

Sensitivity

9.86 pg/mL

Assay Range

25-1600 pg/mL (Cell Culture Supernates, Cell Lysates, Serum, EDTA Plasma, Heparin Plasma)

Product Summary for Human PD-1 QuicKit ELISA

Natural and recombinant human PD-1.

Product Specifications

Measurement

Quantitative ELISA

Detection Method

Colorimetric - 450nm (TMB)

Conjugate

HRP

Reactivity

Human

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.

Sample Values

Serum/Plasma- Samples from apparently healthy volunteers were evaluated for the presence of human PD-1 in this assay. No medical histories were available for the donors used in this study.

Sample TypeMean (pg/mL)Range (pg/mL)Standard Deviation (pg/mL)
Serum (n=10)15180-31481.4
EDTA plasma (n=10)188117-32171.8
Heparin plasma (n=0)188104-37393.1

Cell Culture:

Human peripheral blood mononuclear cells (PBMCs) were cultured at 3 x 106 cells/mL in RPMI supplemented with 10% fetal bovine serum, 2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin and were either left untreated or treated with 10 ug/mL PHA-L and 20 ng/mL of recombinant human (rh) IL-2 for 5 days. After 5 days, cells were then treated with 10 ng/mL PMA and 500 ng/mL Ionomycin for an additional 24 hours before harvesting conditioned media. CD4+ T cells were isolated from PBMCs using MagCellect™ Human CD4+ T Cell Isolation Kit (R&D Systems®, Catalog # MAGH102). For Cloudz™ stimulation, CD4+ T cells were cultured at 5 x 105 /mL using ExCellerate™ Human T Cell Expansion Media, Xeno-Free (R&D Systems, Catalog # CCM030) for the duration. T cells were treated with 25 μL of Cloudz CD3/28 particles per mL of culture media for 5 days. Cell conditioned media was collected by centrifugation and stored at ≤ -20 °C until assayed. After centrifugation, the remaining cells and Cloudz particles were resuspended with 1X Release Buffer. After dissolution of Cloudz particles, cells were lysed as indicated on the next page. 

Precision

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

Inter-Assay Precision (Precision between assays) Two samples of known concentration were tested in ten separate assays to assess inter-assay precision. Assays were performed by at least three technicians.

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

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 1 2
n 20 20 20 20
Mean (pg/mL) 143 871 157 990
Standard Deviation 6.17 19.4 15.0 57.2
CV% 4.3 2.2 9.6 5.8

Recovery for Human PD-1 QuicKit ELISA

The recovery of human PD-1 spiked to three different levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 114 91-131
Cell Lysis Buffer (n=1) 117 94-133
EDTA Plasma (n=2) 104 91-122
Heparin Plasma (n=2) 85 71-103
Serum (n=2) 75 67-82

Linearity

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

Scientific Data Images for Human PD-1 QuicKit ELISA

Human PD-1 ELISA Standard Curve

Human PD-1 QuicKit Natural Linearity Competitor Comparison

Samples containing and/or spiked with high concentrations of PD-1 in various matrices and diluted with appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay. The linearity is between 99%-109% compared to 105%-135% for the top competitor.

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

PD-1 (Programmed Death-1 receptor) is a key regulator of the threshold of immune response and peripheral immune tolerance. It is expressed on activated T cells, B cells, monocytes, and dendritic cells and binds to PD-L1 or PD-L2. PD-1 ligation induces co-inhibitory signals in T cells promoting their apoptosis, anergy, and functional exhaustion. Blockade of the PD-1: PD-L1 interaction using anti-PD-1 antibodies enhances understanding of anti-tumor immunity and advances the potential for cancer immunotherapy.

Long Name

Programmed Death-1

Alternate Names

CD279, PD1, PDCD1, SLEB2

Entrez Gene IDs

5133 (Human); 18566 (Mouse); 301626 (Rat); 100533201 (Porcine); 486213 (Canine); 102123659 (Cynomolgus Monkey)

Gene Symbol

PDCD1

Additional PD-1 Products

Product Documents for Human PD-1 QuicKit ELISA

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human PD-1 QuicKit ELISA

For research use only

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