Human PD-L1 ELISA Kit, Fluorescent (ab229414)
Key features and details
- One-wash 90 minute protocol
- Sensitivity: 2.4 pg/ml
- Range: 2.73 pg/ml - 2800 pg/ml
- Sample type: Cell culture extracts, Cell culture supernatant, Cit plasma, EDTA Plasma, Hep Plasma, Serum, Tissue Extracts, Urine
- Detection method: Fluorescent
- Assay type: Sandwich (quantitative)
- Reacts with: Human
Overview
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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, Hep Plasma, EDTA Plasma, Cit 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% 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 -
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.orgThe 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.
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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
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Storage instructions
Store at +4°C. Please refer to protocols. -
Components 1 x 96 tests 100X Stoplight Red Substrate 1 x 120µl 10X Human PD-L1 Capture Antibody 1 x 600µl 10X Human PD-L1 Detector Antibody (RabMab clone 28-8) 1 x 600µl 10X Wash Buffer PT (ab206977) 1 x 20ml 500X Hydrogen Peroxide (H2O2, 3%) 1 x 50µl 50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml 5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml Antibody Diluent CPI - HAMA Blocker (ab193969) 1 x 6ml Human PD-L1 Lyophilized Recombinant Protein 2 vials Plate Seals 1 unit Sample Diluent NS (ab193972) 1 x 50ml SimpleStep Pre-Coated Black 96-Well Microplate 1 unit Stoplight Red Substrate Buffer 1 x 12ml -
Research areas
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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
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Alternative names
- B7 H
- B7 H1
- B7 homolog 1
see all -
Database links
- Entrez Gene: 29126 Human
- Omim: 605402 Human
- SwissProt: Q9NZQ7 Human
- Unigene: 521989 Human
Images
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Other - Human PD-L1 ELISA Kit, Fluorescent (ab229414)
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.
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Example of human PD-L1 standard curve in Sample Diluent NSBackground-subtracted data values (mean +/- SD) are graphed.
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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 loadThe 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.
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Serum from ten individual healthy human male donors was measured in duplicateInterpolated 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.
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Human peripheral blood mononuclear cells were cultured unstimulated or stimulated with 10 µg/mL PHAConditioned 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.
