Overview

  • Product name
    Human PD-L1 ELISA Kit, Fluorescent
    See all PD-L1 kits
  • Detection method
    Fluorescent
  • Precision
    Intra-assay
    Sample n Mean SD CV%
    Cell Extract 8 5.4%
    Inter-assay
    Sample n Mean SD CV%
    Cell Extract 3 4.1%
  • Sample type
    Cell culture supernatant, Urine, Serum, Cell culture extracts, Tissue Extracts, Heparin Plasma, EDTA Plasma, Citrate Plasma
  • Assay type
    Sandwich (quantitative)
  • Sensitivity
    2.4 pg/ml
  • Range
    2.73 pg/ml - 2800 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%
    Heparin Plasma 92 89% - 93%
    EDTA Plasma 85 81% - 88%
    Citrate Plasma 84 79% - 88%

  • Assay time
    1h 30m
  • Assay duration
    One step assay
  • Species reactivity
    Reacts with: Human
    Does not react with: Mouse, Rat, Cow
  • Product overview

    PD-L1 in vitro CatchPoint SimpleStep ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of PD-L1 protein in human serum, plasma, cell culture supernatant, urine, and cell and tissue extracts.


    This CatchPoint SimpleStep ELISA kit has been optimized for Molecular Devices Microplate Readers. Click here for a list of recommended Microplate Readers.
    If using a Molecular Devices’ plate reader supported by SoftMax® Pro software, a preconfigured protocol for these CatchPoint SimpleStep ELISA Kits is available with all the protocol and analysis settings at www.softmaxpro.org


    The CatchPoint SimpleStep ELISA employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material.  CatchPoint HRP Development Solution containing the Stoplight Red Substrate is added. During incubation, the substrate is catalyzed by HRP generating a fluorescent product. Signal is generated proportionally to the amount of bound analyte and the intensity is measured in a fluorescence plater reader at 530/570/590 nm Excitation/Cutoff/Emission. 


     

  • 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.

     

  • Tested applications
    Suitable for: Sandwich ELISAmore details
  • 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 Human PD-L1 Capture Antibody 1 x 600µl
    10X Human PD-L1 Detector Antibody (RabMab clone 28-8) 10X (205 1 x 600µl
    Human PD-L1 Lyophilized Recombinant Protein 2 vials
    Antibody Diluent CPI 1 x 6ml
    10X Wash Buffer PT (ab206977) 1 x 20ml
    5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml
    50X Cell Extraction Enhancer Solution 1 x 1ml
    Stoplight Red Substrate Buffer 1 x 12ml
    100X Stoplight Red Substrate 1 x 120µl
    500X Hydrogen Peroxide (H2O2, 3%) 1 x 50µl
    Sample Diluent NS (ab193972) 1 x 50ml
    SimpleStep Pre-Coated Black 96-Well Microplate 1 unit
    Plate Seals 1 unit
  • 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

Applications

Our Abpromise guarantee covers the use of ab229414 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Sandwich ELISA Use at an assay dependent concentration.

Images

  • ELISA Protocol Summary
  • Background-subtracted data values (mean +/- SD) are graphed.

  • 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.

  • 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.

  • 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.

Protocols

References

ab229414 has not yet been referenced specifically in any publications.

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