Astana Biomed Group, an authorized Abcam distributor in Central Asia

Human PD-L1 ELISA Kit [28-8] (ab214565)

Key features and details

  • One-wash 90 minute protocol
  • Sensitivity: 2.91 pg/ml
  • Range: 21.87 pg/ml - 1400 pg/ml
  • Sample type: Cell culture extracts, Cell culture supernatant, Plasma, Serum, Tissue Extracts, Urine
  • Detection method: Colorimetric
  • Assay type: Sandwich (quantitative)
  • Reacts with: Human

Overview

  • Product name

    Human PD-L1 ELISA Kit [28-8]
    See all PD-L1 kits

  • Detection method

    Colorimetric

  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Overall 8 5.4%
    Inter-assay
    Sample n Mean SD CV%
    Overall 3 4.1%

  • Sample type

    Cell culture supernatant, Urine, Serum, Plasma, Cell culture extracts, Tissue Extracts

  • Assay type

    Sandwich (quantitative)

  • Sensitivity

    2.91 pg/ml

  • Range

    21.87 pg/ml - 1400 pg/ml

  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Cell culture supernatant 103 102% - 105%
    Urine 91 86% - 96%
    Serum 86 82% - 91%
    Cell culture extracts 102 100% - 103%
    Tissue Extracts 105 104% - 107%
    Hep Plasma 92 89% - 93%
    EDTA Plasma 85 81% - 88%
    Cit plasma 84 79% - 88%

  • Assay time

    1h 30m

  • Assay duration

    One step assay

  • Species reactivity

    Reacts with: Human
    Does not react with: Cow, Cynomolgus monkey, Macaque monkey

  • Product overview

    Human PD-L1 ELISA Kit [28-8] (ab214565) has been re-developed with a new recombinant antibody to provide improved consistency and security of supply. This version, ab214565, will be discontinued in July 2021.


    The new version is available as ab277712. It retains the recombinant PD-L1 antibody clone [28-8].


    Human PD-L1 ELISA Kit [28-8] (ab214565) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of PD-L1 [28-8] protein in cell culture extracts, cell culture supernatant, serum, tissue extracts, urine, and plasma. It uses our proprietary SimpleStep ELISA® technology. Quantitate Human PD-L1 [28-8] with 2.91 pg/ml sensitivity.


    SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:


            - Single-wash protocol reduces assay time to 90 minutes or less
            - High sensitivity, specificity and reproducibility from superior antibodies
            - Fully validated in biological samples
            - 96-wells plate breakable into 12 x 8 wells strips


    A 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.


    ASSAY SPECIFICITY


    This kit recognizes both native and recombinant human PD-L1 protein in human serum, plasma, and cell culture supernatant, cell and tissue extract samples only.


    CROSS REACTIVITY


    Recombinant mouse PD-L1 was prepared at 50 ng/mL and 1250 pg/mL and assayed for cross reactivity. No cross-reactivity was observed.


    INTERFERENCE


    Recombinant PD-1 was prepared at 50 ng/mL and 1250 pg/mL and tested for interference. No interference with was observed.


    SPECIES REACTIVITY


    This kit recognizes human PD-L1 protein.


    Other species reactivity was determined by measuring 50% serum samples of various species, interpolating the protein concentrations from the human standard curve, and expressing the interpolated concentrations as a percentage of the protein concentration in human serum assayed at the same dilution.


     Reactivity

     

  • Notes

    PD-L1 (also known as CD274 or B7-H1) is a membrane bound glycoprotein involved in regulation of the immune system. PD-L1 is expressed on a variety of inflammatory-activated cells as well as some carcinomas and in melanoma. PD-L1 binds to PD-1 and CD80, where it can suppress T cell activation and proliferation as well as induce apoptosis. Levels of PD-L1 are increased in the plasma of cancer patients as well as in cerebrospinal fluid of gliomas. PD-L1 can bind PD-1 in order to regulate T cell apoptosis.

    Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
    It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

  • Platform

    Pre-coated microplate (12 x 8 well strips)

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Wash Buffer PT (ab206977) 1 x 20ml
    5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml
    Antibody Diluent CPI - HAMA Blocker (ab193969) 1 x 6ml
    10X Human PD-L1 Capture Antibody 1 x 600µl
    Human PD-L1 Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    10X Human PD-L1 Detector Antibody (RabMab clone 28-8) 1 x 600µl
    Sample Diluent NS (ab193972) 1 x 50ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml

  • Research areas

  • Function

    Involved in the costimulatory signal, essential for T-cell proliferation and production of IL10 and IFNG, in an IL2-dependent and a PDCD1-independent manner. Interaction with PDCD1 inhibits T-cell proliferation and cytokine production.

  • Tissue specificity

    Highly expressed in the heart, skeletal muscle, placenta and lung. Weakly expressed in the thymus, spleen, kidney and liver. Expressed on activated T- and B-cells, dendritic cells, keratinocytes and monocytes.

  • Sequence similarities

    Belongs to the immunoglobulin superfamily. BTN/MOG family.
    Contains 1 Ig-like C2-type (immunoglobulin-like) domain.
    Contains 1 Ig-like V-type (immunoglobulin-like) domain.

  • Cellular localization

    Cell membrane and Endomembrane system.
  • Information by UniProt

  • Alternative names

    • B7 H
    • B7 H1
    • B7 homolog 1
    • B7-H1
    • B7H
    • B7H1
    • CD 274
    • CD274
    • CD274 antigen
    • CD274 molecule
    • MGC142294
    • MGC142296
    • OTTHUMP00000021029
    • PD L1
    • PD-L1
    • PD1L1_HUMAN
    • PDCD1 ligand 1
    • PDCD1L1
    • PDCD1LG1
    • PDL 1
    • PDL1
    • Programmed cell death 1 ligand 1
    • Programmed death ligand 1
    • RGD1566211
    see all

  • Database links

    Images

    • Other - Human PD-L1 ELISA Kit [28-8] (ab214565)
      Other - Human PD-L1 ELISA Kit [28-8] (ab214565)

      SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

       

    • Interpolated concentrations of native PD-L1 in human Jurkat stimulated with LPS and IFN-gamma, placenta and thyroid based on a 1,000 µg/mL extract load.
      Interpolated concentrations of native PD-L1 in human Jurkat stimulated with LPS and IFN-gamma, placenta and thyroid based on a 1,000 µg/mL extract load.

      The concentrations of PD-L1 were measured in duplicate and interpolated from the PD-L1 standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean PD-L1 concentration was determined to be 404.3 pg/mL in Jurkat stimulated with LPS and IFN-gamma, 628.4 pg/mL in placenta and 207.5 pg/mL in thyroid extracts.

    • Serum from ten individual healthy human male donors was measured in duplicate.
      Serum from ten individual healthy human male donors was measured in duplicate.

      Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).  PD-L1 was measured in 6 donor serum samples and the remaining 4 samples measured less than the lowest point of the PD-L1 standard curve. Of those measured, the mean PD-L1 concentration was determined to be 44.5 pg/mL with a range of 34.4 – 75.3 pg/mL.

    • Example of human PD-L1 standard curve in 1X Cell Extraction Buffer PTR.
      Example of human PD-L1 standard curve in 1X Cell Extraction Buffer PTR.

      Background-subtracted data values (mean +/- SD) are graphed.

    • Example of human PD-L1 standard curve in 1X Cell Extraction Buffer PTR.
      Example of human PD-L1 standard curve in 1X Cell Extraction Buffer PTR.

      The PD-L1 standard curve was prepared as described. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.

    • Example of human PD-L1 standard curve in Sample Diluent NS.
      Example of human PD-L1 standard curve in Sample Diluent NS.

      Background-subtracted data values (mean +/- SD) are graphed.

    • Example of human PD-L1 standard curve in Sample Diluent NS.
      Example of human PD-L1 standard curve in Sample Diluent NS.

      The PD-L1 standard curve was prepared as described. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.

    • Human peripheral blood mononuclear cells were cultured unstimulated or stimulated with 10 µg/mL PHA.
      Human peripheral blood mononuclear cells were cultured unstimulated or stimulated with 10 µg/mL PHA.

      Conditioned media was harvested after 48 hours. PD-L1 was measured in 100% unstimulated and PHA stimulated PBMC supernatant. The concentrations of PD-L1 were measured in duplicate and interpolated from the PD-L1 standard curves. The interpolated values are plotted (mean +/- SD, n=2). The mean PD-L1 concentration was determined to be 59.7 pg/mL in PHA stimulated PBMC supernatant. There was no detectable signal in unstimulated supernatant.

    • Linearity of dilution.
      Linearity of dilution.

      Linearity of dilution is determined based on interpolated values from the standard curve. Linearity of dilution defines a sample concentration interval in which interpolated target concentrations are directly proportional to sample dilution.

      Native PD-L1 was measured in the following biological samples in a 2-fold dilution series. Sample dilutions are made in 1X Cell Extraction Buffer PTR.

    • Linearity of dilution.
      Linearity of dilution.

      Linearity of dilution is determined based on interpolated values from the standard curve. Linearity of dilution defines a sample concentration interval in which interpolated target concentrations are directly proportional to sample dilution.

      Recombinant PD-L1 was spiked into the following biological samples and diluted in a 2-fold dilution series in Sample Diluent NS.

    • Linearity of dilution.
      Linearity of dilution.

      Linearity of dilution is determined based on interpolated values from the standard curve. Linearity of dilution defines a sample concentration interval in which interpolated target concentrations are directly proportional to sample dilution.

      Recombinant PD-L1 was spiked into the following biological sample and diluted in a 2-fold dilution series in Sample Diluent NS.

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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