SimpleStep

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

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Other - Human PD-L1 ELISA Kit [28-8] (ab214565)
  • 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.
  • Serum from ten individual healthy human male donors was measured in duplicate.
  • 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.
  • Example of human PD-L1 standard curve in Sample Diluent NS.
  • Example of human PD-L1 standard curve in Sample Diluent NS.
  • Human peripheral blood mononuclear cells were cultured unstimulated or stimulated with 10 µg/mL PHA.
  • Linearity of dilution.
  • Linearity of dilution.
  • Linearity of dilution.

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

製品の概要

  • 製品名

    Human PD-L1 ELISA Kit [28-8]
    PD-L1 キット 製品一覧

  • 検出方法

    Colorimetric

  • 再現性

    Intra-Assay（同時再現性）
    サンプル N 平均値 SD CV%
    Overall 8 5.4%
    Inter-Assay（日差再現性）
    サンプル N 平均値 SD CV%
    Overall 3 4.1%

  • サンプルの種類

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

  • アッセイタイプ

    Sandwich (quantitative)

  • 検出感度

    2.91 pg/ml

  • 検出範囲

    21.87 pg/ml - 1400 pg/ml

  • 添加回収試験

    特定サンプルでの回収試験
    サンプルの種類 平均 % 測定範囲
    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%

  • 全工程の試験時間

    1h 30m

  • ステップ

    One step assay

  • 種交差性

    交差種: Human
    非交差種: Cow, Cynomolgus monkey, Macaque monkey

  • 製品の概要

    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 < 3% was determined for the following species: Mouse, Rat, Cow


     

  • 特記事項

    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.

  • 試験プラットフォーム

    Pre-coated microplate (12 x 8 well strips)

製品の特性

  • 保存方法

    Store at +4°C. Please refer to protocols.
  • 内容 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

  • 研究分野

  • 機能

    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.

  • 組織特異性

    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.

  • 配列類似性

    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.

  • 細胞内局在

    Cell membrane and Endomembrane system.
  • Information by UniProt

  • 別名

    • B7 H
    • B7 H1
    • B7 homolog 1
    • B7-H1
    • B7H
    • B7H1
    • CD 274
    • 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

  • 参照データベース

    画像

    • 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.

    参考文献 (16)

    ab214565 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

    ab214565 は 16 報の論文で使用されています。

    • Lu T  et al. High Soluble Programmed Death-Ligand 1 Predicts Poor Prognosis in Patients with Nasopharyngeal Carcinoma. Onco Targets Ther 13:1757-1765 (2020). PubMed: 32161471
    • Asanuma K  et al. Soluble programmed death-ligand 1 rather than PD-L1 on tumor cells effectively predicts metastasis and prognosis in soft tissue sarcomas. Sci Rep 10:9077 (2020). PubMed: 32493964
    • Abu Hejleh T  et al. The clinical significance of soluble PD-1 and PD-L1 in lung cancer. Crit Rev Oncol Hematol 143:148-152 (2019). PubMed: 31675543
    • Azadi S  et al. Upregulation of PD-L1 expression in breast cancer cells through the formation of 3D multicellular cancer aggregates under different chemical and mechanical conditions. Biochim Biophys Acta Mol Cell Res 1866:118526 (2019). PubMed: 31398408
    • Gong B  et al. Secreted PD-L1 variants mediate resistance to PD-L1 blockade therapy in non-small cell lung cancer. J Exp Med 216:982-1000 (2019). PubMed: 30872362
    View all Publications for this product

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    PD-L1 quantification in HCT116 colon cancer cells

     Excellent Excellent 5/5 (Ease of Use)
    Abreviews
    abreview image
    We quantified the PD-L1 concentration in cell supernatants from HCT116 colon cancer cells treated with pure anthocyanins and extracts from black lentils, as well as commercial drugs (ATZPMB). We used this kit for obtaining precise and accurate values of the PD-L1 concentration since this protein is essential in the immuno-modulation of cancer, as we were targeting the use of these extracts as immuno-therapy.

    Abcam user community

    Verified customer

    投稿 Jun 06 2019

    Question

    I see that the "sensitivity" of this kit is 2.91 pg/ml. Does this number represent the lower limit of detection (LLOD) or the lower limit of quantification (LLOQ)?

    Read More
    Answer

    We refer to sensitivity as the "Minimal Detectable Dose" (MDD) which is defined as follows on page 19 of the protocol booklet linked to the https://www.abcam.com/human-pd-l1-elisa-kit-28-8-ab214565.html near the upper right.


    From page 19 of the online protocol: The MDD was determined by calculating the mean of zero standard replicates and adding 2 standard deviations, then extrapolating the corresponding concentration.


    It is the lower limit of detection. My understanding is that the lower limit of quantification is the value of the lowest standard, ˜22pg/mL. Please let me know if you have any other questions.

    Read More

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