Home / PD-1 / Human PD-1 DuoSet ELISA

Human PD-1 DuoSet ELISA

Catalog #: DY1086
11 Citations Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
DY1086
Ancillary Products Available
DY008B: DuoSet ELISA Ancillary Reagent Kit 2
DY999B: Substrate Reagent Pack
DY999: Substrate Reagent Pack
Human PD-1 ELISA Standard Curve
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Product Details
Procedure
Citations (11)
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Summary Product Features Kit Content Other Reagents Required Data Examples Product Datasheets Preparation and Storage Background

Human PD-1 DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
Assay Range
156.0 - 10,000 pg/mL
Sufficient Materials
For fifteen 96-well plates*
Specificity
Please see the product datasheet

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human PD-1. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet ELISA.

Product Features

  • Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
  • Development protocols are provided to guide further assay optimization
  • Assay can be customized to your specific needs
  • Economical alternative to complete kits

Kit Content

  • Capture Antibody
  • Detection Antibody
  • Recombinant Standard
  • Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Other Reagents Required


PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or equivalent

Reagent Diluent: 1% BSA in PBS, pH 7.2-7.4, 0.2 m filtered (R&D Systems Catalog # DY995). Quality of BSA is critical (see Technical Hints).

Blocking Buffer: 1% BSA in PBS, pH 7.2-7.4, 0.2 m filtered (R&D Systems Catalog # DY995). Quality of BSA is critical (see Technical Hints).

Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H2SO4 (Catalog # DY994)

Microplates: R&D Systems (Catalog # DY990), or equivalent

Plate Sealers: ELISA Plate Sealers (Catalog # DY992), or equivalent

 

Scientific Data

N/A Human PD-1 ELISA Standard Curve View Larger

Human PD-1 ELISA Standard Curve

Product Datasheets

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Product Datasheet
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SDS

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
Entrez Gene IDs:
5133 (Human); 18566 (Mouse); 301626 (Rat); 100533201 (Porcine); 486213 (Canine); 102123659 (Cynomolgus Monkey)
Alternate Names:
CD279 antigen; CD279; hPD-1; PD1; PD-1; PD1hPD-l; PDCD1; programmed cell death 1; programmed cell death protein 1; Protein PD-1; SLEB2

Assay Procedure

GENERAL ELISA PROTOCOL

Plate Preparation

  1. Dilute the Capture Antibody (to the working concentration stated in the product datasheet ) in PBS without carrier protein. Immediately coat a 96-well microplate with 100 µL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
  2. Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 µL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
  3. Block each well of the microplate as recommended in the product datasheet. Incubate at room temperature for a minimum of 1 hour.

    Note: The recommended Reagent Diluent typically contains 1% BSA. Some DuoSet Development Kits require alternative blocking agents, or for plates to be blocked overnight with a higher percentage of BSA, please see the product datasheet for details.
     
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

 

PRECAUTION
The Stop Solution suggested for use with this kit is an acid solution. Wear eye, hand, face and clothing protection when using this material.

Assay Procedure

  1. Add 100 µL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. Add 100 µL of the Detection Antibody, diluted in Reagent Diluent (as recommended in the product datasheet), to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. Add 100 µL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 µL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 µL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.

Citations for Human PD-1 DuoSet ELISA

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

11 Citations: Showing 1 - 10
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  1. Robust Preanalytical Performance of Soluble PD-1, PD-L1 and PD-L2 Assessed by Sensitive ELISAs in Blood
    Authors: K Krueger, Z Mayer, M Kottmaier, M Gerckens, S Boeck, P Luppa, S Holdenried
    Biomedicines, 2022-10-11;10(10):.
    Species: Human
    Sample Types: Plasma
  2. High Quality Performance of Novel Immunoassays for the Sensitive Quantification of Soluble PD-1, PD-L1 and PD-L2 in Blood
    Authors: K Krueger, Z Mayer, M Gerckens, S Boeck, P Luppa, S Holdenried
    Biomedicines, 2022-09-26;10(10):.
    Species: Human
    Sample Types: Plasma
  3. High Levels of Soluble Programmed Death-1 Are Associated with Virological Response in Chronic Hepatitis B patients after Antiviral Treatment
    Authors: N Tan, H Luo, Q Kang, J Pan, R Cheng, H Xi, H Chen, Y Han, Y Yang, X Xu
    Virus research, 2021-12-18;0(0):198660.
    Species: Human
    Sample Types: Serum
  4. Increased Plasma Soluble PD-1 Concentration Correlates with Disease Progression in Patients with Cancer Treated with Anti-PD-1 Antibodies
    Authors: R Ohkuma, K Ieguchi, M Watanabe, D Takayanagi, T Goshima, R Onoue, K Hamada, Y Kubota, A Horiike, T Ishiguro, Y Hirasawa, H Ariizumi, J Tsurutani, K Yoshimura, M Tsuji, Y Kiuchi, S Kobayashi, T Tsunoda, S Wada
    Biomedicines, 2021-12-16;9(12):.
    Species: Human
    Sample Types: Plasma
  5. Soluble PD-L1 Is an Independent Prognostic Factor in Clear Cell Renal Cell Carcinoma
    Authors: G Larrinaga, JD Solano-Itu, P Errarte, M Unda, A Loizaga-Ir, A Pérez-Fern, E Echevarría, A Asumendi, C Manini, JC Angulo, JI López
    Cancers, 2021-02-07;13(4):.
    Species: Human
    Sample Types: Plasma
  6. Soluble PD-1 but Not PD-L1 Levels Predict Poor Outcome in Patients with High-Risk Diffuse Large B-Cell Lymphoma
    Authors: H Vajavaara, JB Mortensen, SK Leivonen, IM Hansen, M Ludvigsen, H Holte, J Jørgensen, M Bjerre, F d'Amore, S Leppä
    Cancers, 2021-01-22;13(3):.
    Species: Human
    Sample Types: Serum
  7. Characterization of Human NK Cell-Derived Exosomes: Role of DNAM1 Receptor In Exosome-Mediated Cytotoxicity Against Tumor
    Authors: ALD Pace, N Tumino, F Besi, C Alicata, LA Conti, E Munari, E Maggi, P Vacca, L Moretta
    Cancers (Basel), 2020-03-12;12(3):.
    Species: Human
    Sample Types: Cell Culture Lysates
  8. Immune checkpoint inhibitor PD-1 pathway is down-regulated in synovium at various stages of rheumatoid arthritis disease progression
    Authors: Y Guo, AM Walsh, M Canavan, MD Wechalekar, S Cole, X Yin, B Scott, M Loza, C Orr, T McGarry, M Bombardier, F Humby, SM Proudman, C Pitzalis, MD Smith, JR Friedman, I Anderson, L Madakamuti, DJ Veale, U Fearon, S Nagpal
    PLoS ONE, 2018-02-28;13(2):e0192704.
    Species: Human
    Sample Types: Serum
  9. Serum levels of soluble programmed death protein 1 (sPD-1) and soluble programmed death ligand 1 (sPD-L1) in advanced pancreatic cancer
    Authors: S Kruger, ML Legenstein, V Rösgen, M Haas, DP Modest, CB Westphalen, S Ormanns, T Kirchner, V Heinemann, S Holdenried, S Boeck
    Oncoimmunology, 2017-03-31;6(5):e1310358.
    Species: Human
    Sample Types: Serum
  10. Tumor localized secretion of soluble PD1 enhances oncolytic virotherapy
    Authors: MY Bartee, KM Dunlap, E Bartee
    Cancer Res, 2017-03-17;0(0):.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  11. Circulating programmed death-1 as a marker for sustained high hepatitis B viral load and risk of hepatocellular carcinoma.
    Authors: Cheng H, Kang P, Chuang Y, Wang Y, Jan M, Wu C, Lin C, Liu C, Liaw Y, Lin S, Chen P, Lee S, Yu M
    PLoS ONE, 2014-11-26;9(11):e95870.
    Species: Human
    Sample Types: Plasma

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