PerkinElmer
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PD-L1 (human) AlphaLISA Detection Kit, 500 assay points

The AlphaLISA® Human PD-L1 Detection Kit is designed for detection and quantitation of human Programmed death ligand 1 (PD-L1; CD274/B7-H1) in cell culture media or serum using a homogeneous (no-wash steps, no separation steps) assay.

For research use only; not for diagnostic procedures. All products to be used in accordance with applicable laws and regulations including without limitation, consumption & disposal requirements under European REACH regulations (EC 1907/2006).

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Unit Size
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AL355HV
100 assay points
828.00 USD
 
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AL355C
500 assay points
1758.00 USD
 
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AL355F
5,000 assay points
11600.00 USD
 
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Overview

Formats:

  • Our HV (100 assay point) kits allow you to run 100 wells in 96-well format, using a 100 µL reaction volume (10 µL of sample).
  • Our 500 assay point kit allows you to run 500 wells in 96-well or 384-well format, using a 50 µL reaction volume (5 µL of sample).
  • Our 5,000 assay point kit allows you to run 5,000 wells in 96-well or 384-well format, using a 50 µL reaction volume (5 µL of sample).

Features:

  • No-wash steps, no separation steps
  • ELISA alternative technology
  • Sensitive detection
  • Broad sample compatibility
  • Small sample volume
  • Results in less than 3 hours
  • Half the time of an ELISA assay

Programmed death ligand 1 (PD-L1), also known as cluster of differentiation 274 (CD274) or B7 homolog1 (B7-H1) belongs to the growing B7 family of immune proteins and has been demonstrated to play a role in the regulation of immune responses and peripheral tolerance. Human PD-L1 is constitutively expressed in several organs such as heart, skeletal muscle, placenta and lung, and in lower amounts in thymus, spleen, kidney and liver. PD-L1, together with PD-L2, are two ligands for PD-1 (programmed death 1), a member of the CD28 family of immunoreceptors. By binding to PD-1 on activated T-cells and B-cells, PD-L1 may inhibit ongoing T-cell responses by inducing apoptosis and arresting cell-cycle progression. Accordingly, it leads to growth of immunogenic tumor growth by increasing apoptosis of antigen specific T cells and may contribute to immune evasion by cancers. PD-L1 thus is regarded as promising therapeutic target for human autoimmune disease and malignant cancers.

AlphaLISA technology allows the detection of molecules of interest in a no-wash, highly sensitive, quantitative assay. In an AlphaLISA assay, a biotinylated anti-analyte antibody binds to the Streptavidin-coated Donor beads while another anti-analyte antibody is conjugated to AlphaLISA Acceptor beads. In the presence of the analyte, the beads come into close proximity. The excitation of the Donor beads causes the release of singlet oxygen molecules that triggers a cascade of energy transfer in the Acceptor beads, resulting in a sharp peak of light emission at 615 nm.

Specifications

Assay Target PD-L1
Assay Target Class Protein
Automation Compatible Yes
Detection Method Alpha
Experimental Type In vitro
Product Brand Name AlphaLISA
Shipping Condition Blue Ice
Therapeutic Area Cancer
Unit Size 500 assay points
Resources, Events & More
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Application Brief

Eight Limitations of ELISA and How to Overcome Them Using Alternative Technologies

The introduction of enzyme-linked immunosorbent assays (ELISAs) in the early 1970’s offered researchers a non-radiometric immunoassay platform without compromising sensitivity. Over the last 50 years scientists have made huge strides in disease research and drug discovery and a demand for greater assay throughput and sensitivity has evolved. In response, more robust immunoassays have been developed to address some of the limitations of the standard, colorimetric ELISA.

Find out about the most common limitations of traditional ELISAs and how different ELISA alternative technologies address these limitations.

PDF 1 MB

Application Note

Measuring PD-L1 Expression in Breast Cancer Cell Lines with AlphaLISA

Too many candidates, too little time. The lack of robust, rapid, high-throughput assays to identify and qualify potential therapeutic targets in areas such as cancer research continues to cost valuable time. What if you could increase assay throughput without compromising sensitivity, obtain more data points from each sample and eliminate tedious wash steps? Find out how AlphaLISA® assay technology, combined with the EnVision® multimode plate reader, provides a fast, powerful, homogeneous platform for screening potential inhibitors of PD-L1 (a protein associated with breast cancer tumor cells) expression in human cells.

PDF 4 MB
Quantifying Changes in CD28 and CTLA-4 Levels in Peripheral Blood Mononuclear Cells with AlphaLISA Technology

One approach to immunotherapy is the modulation of immune checkpoints that are critical in regulating the degree and duration of immune system responses and preventing autoimmunity.

In this application note, you will learn:

  • Why AlphaLISA® technology is an effective assay for the detection and quantification of biomarkers such as CD28 and CTLA-4
  • How to run an AlphaLISA assay to detect and quantify immuno-oncology biomarkers
  • Benefits of using Alpha technology for biomarker quantification assays
  • An example of the data and analysis you can generate for biomarker assays

PDF 1 MB
Rapid, No-wash Measurement of Immune Checkpoint Molecules and Cytokines in Co-cultures of Immune Cells and Cancer Cell Lines

Breast cancer tumors can adapt to immune cell infiltration by responding to the increased concentration of interferon gamma (IFN-ɣ) and other cytokines secreted by subsets of T lymphocytes with the upregulation of the immune checkpoint proteins such as Programmed cell death ligand 1 (PD-L1). These checkpoint proteins allow the tumors to evade immune targeting and reduce the immune response, thus promoting tumor progression.

In this application note, you will learn:

  • How exogenous addition of IFN-ɣ effects the expression of PD-L1 and secretion of several cytokines in cultures of HCC38 cells (a triple-negative breast cancer cell line)
  • How co-culturing activated immune cells and breast cancer cells stimulates differential expression of some immune checkpoint and inflammatory biomarkers compared to culturing cells alone with PBMC-conditioned media
  • How to rapidly measure multiple biomarkers in cell culture supernatant and lysates from the same wells of a culture dish to examine protein expression profiles from cancer and immune cell culture models using AlphaLISA® assays together with ATPlite 1step and the EnVision® multimode plate reader.

PDF 2 MB
Using AlphaLISA Biomarker Kits to Assess Effects of PBMC-Conditioned Media on 3D and 2D Cell Culture Models of Breast Cancer

Various cytokines are secreted during an active immune response that can have modulatory effects on target cell populations, including interferon gamma (IFN-ɣ), tumor necrosis factor alpha (TNFa) and several interleukins.

In this application note, you will learn how we investigated:

  • The effects of secreted factors on a triple-negative breast cancer cell line by treatment with conditioned media from activated PBMCs
  • Whether the modulatory effects of cytokines secreted by infiltrating immune is different when studied in 3D cell cultures
  • How to detect and quantify multiple immune-modulated proteins from the same wells in complex cell models grown in 3D spheroid microplates and traditional 2D cell culture using AlphaLISA® technology and EnVision® multimode plate reader in a workflow with ATPlite 1step and Opera® Phenix high-content screening system

PDF 2 MB

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