PerkinElmer
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IL-1β (human) AlphaLISA Detection Kit, 500 Assay Points

The AlphaLISA® Human Interleukin-1β (IL1β) Detection Kit is designed for detection and quantitation of human IL-1β in serum, buffered solution or cell culture medium in 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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AL220C
500 assay points
1811.00 USD
 
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AL220F
5,000 assay points
12000.00 USD
 
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Overview

Formats:

  • 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

IL1β and IL1β are central players of the immune response, displaying roles in inflammation both at local and systemic levels. Despite they seem to display very similar functions, these proteins are encoded by two independent genes sharing only ~30% identity. IL1β is synthesized as a 31 kDa precursor that is cleaved by Caspase-1 (ICE) into the active 17 kDa form, and eventually released into the extracellular space. Its production has been reported in many cell types including brain and, importantly, monocytic and periferal blood mononuclear cells. After binding to its receptor, IL-1RI, IL1β triggers a cascade of kinase signaling pathways that lead to the activation of transcription factors like NFκB and AP-1, eventually activating the expression of genes such as MIP-2 and C-reactive protein.

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 IL1β
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 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

Brochure

Data Sheet

Poster

All-In-One-Well AlphaLISA Assays for Direct Biomarker Quantification in Cell Cultures

Biomarker levels were measured directly in CulturPlates-96 and-384 in a simple, fast, all-in-one-well AlphaLISA assay format. The elimination of transfer and wash steps simplifies cellular assays, reduces variability and significantly reduces hands-on time and costs associated with consumables. Integral plasma membrane (EGFR) and secreted solubleproteins (TNFa, IL1ß, IL6, IL8) were successfully determined, on adherent or suspension cells, using the standard AlphaLISA Immunoassay buffer.

PDF 288 KB
AlphaLISA Assays are Homogeneous Sensitive Immunoassays for Detection of Analytesin a Variety of Biological Matrices

The AlphaLISA® assay is a homogeneous immunoassay alternative to classical ELISA. AlphaLISA assays were originally utilized to detect analytesin cell cultures upernatants or serum/plasma samples.

PDF 273 KB
Development of New AlphaLISA No-wash Immunoassay Kits for Sensitive, Rapid and Efficient Quantification of Cytokines

Enzyme-linked Immunosorbent Assay is the most widely Kits adopted method for detection and quantification of cytokines and other biomarkers. This traditional technology offers good,selectivity, sensitivity and assay versatility; however, it has certain disadvantages such as limited dynamic range and low throughput due to the numerous wash steps.

PDF 279 KB