PerkinElmer
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IFN-γ (human) AlphaLISA Biotin-Free Detection Kit, 500 Assay Points

The AlphaLISA Human IFNγ Biotin-Free Detection Kit is designed for the quantitative detection of human IFNγ in serum, cell culture medium, and other samples types using a homogeneous (no wash steps, no separation steps) assay. The biotin-free kit uses anti-DIG (anti-Digoxin) Donor beads instead of streptavidin Donor beads, which makes the kit compatible with high-biotin culture media and other sample types that contain high levels biotin (including brain/liver tissue extracts, milk and eggs).

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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AL327HV
100 assay points
828.00 USD
 
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AL327C
500 assay points
1811.00 USD
 
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AL327F
5,000 assay points
12000.00 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 and Benefits:

  • No-wash steps, no separation steps
  • Compatible with high biotin samples, including RPMI and high-biotin tissue extracts
  • ELISA alternative technology
  • Sensitive detection
  • Broad sample compatibility
  • Results in less than 3 hours

Interferons (IFNs) activity has been discovered due to their antiviral effects. In humans, there are three families of IFNs: IFN type I (IFN-α, β, ω, ε, and κ), IFN type II (one single representative, IFN-γ), and IFN type III (IFN-λ1-3). Antigens and mitogens stimulate in Natural Killer (NK) and activated helper T lymphocytes (Th1) the production of IFN-γ. Human IFN-γ is a 140 amino acids polypeptide that shows multiple effects; it induces the production of cytokines, upregulates the expression of class I and II MHC antigens, and leukocyte adhesion molecules. It also activates macrophages, enhances the secretion of immunoglobulins by B cells, and potentiates Th1 cell expansion. Response to IFN-γ is mediated by the heterodimeric IFN-γ Receptor, triggering a signaling cascade involving JAK1, JAK2, and STAT1. Importantly, IFNs have been proved to be effective in the treatment of several viral infections and cancers.

AlphaLISA technology allows the detection of molecules of interest in a no-wash, highly sensitive, quantitative assay. In an AlphaLISA biotin-free assay, a DIG-labeled anti-analyte antibody binds to the anti-DIG-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 IFNγ
Assay Target Class Cytokine
Automation Compatible Yes
Detection Method Alpha
Experimental Type In vitro
Product Brand Name AlphaLISA
Shipping Condition Blue Ice
Therapeutic Area Inflammation
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

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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