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AlphaLISA Porcine IL-12 p70 Detection Kit, 100 Assay Points

The AlphaLISA™ immunoassay kit for porcine IL-12 enables the quantitative determination of porcine interleukin-12 in serum, buffered solution, and cell culture supernatants using a homogeneous AlphaLISA assay (no wash steps).

Feature Specification
Application Protein Quantification
Dynamic Range 5 - 100,000 pg/mL  
Limit of Detection 5 pg/mL  
Limit of Quantification 16 pg/mL  
Sample Volume 10 µL

The AlphaLISA™ immunoassay kit for porcine IL-12 enables the quantitative determination of porcine interleukin-12 in serum, buffered solution, and cell culture supernatants using a homogeneous AlphaLISA assay (no wash steps).

Product variants
Unit Size: 100 assay points
Part #:
AL554HV
List price
USD 972.42
Your online price:
Unit Size: 500 assay points
Part #:
AL554C
List price
USD 4,208.58
Your online price:
Unit Size: 5,000 assay points
Part #:
AL554F
List price
USD 29,066.60
Your online price:
For research use only. Not for use in diagnostic procedures. All products to be used in accordance with applicable laws and regulations including without limitation, consumption, and disposal requirements under European REACH regulations (EC 1907/2006).

Overview

Formats:

  • Our 100 assay point kit allows 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

Interleukin 12 (IL-12) belongs to IL-12 family consisting IL-12, IL-23, IL-27, IL-35. IL-12 is encoded by two separate genes, IL-12A (p35) and IL-12B (p40). The active heterodimer (referred to as p70 or IL-12 p70) and a homodimer of p40 are formed following protein synthesis. In response to antigenic stimulation, IL12 proteins are produced by dendritic cells, macrophages, neutrophils, and B-cells. It is known to stimulate the production of interferon-gamma (IFN-γ) and tumor necrosis factor-alpha (TNF-α) from T cells and natural killer (NK) cells, while reducing IL-4 mediated suppression of IFN-γ. IL12 is thought to be one of the cytokines that plays an essential role in inflammatory diseases such as rheumatoid arthritis, allograft rejection, and atherosclerosis. This kit is designed to detect Interleukin 12 (IL-12) p70 as an active heterodimer in procine serum, plasma, and cell culture supernatants.

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.

How it works

Principle of the AlphaLISA assay

The AlphaLISA assay is based on an AlphaLISA sandwich immunoassay involving a biotinylated anti-analyte antibody bound to Streptavidin-coated AlphaLISA Donor beads and an anti-analyte antibody conjugated to AlphaLISA Acceptor beads. Both antibodies are directed against the analyte of interest. In the presence of the target, both antibodies bind to analyte and the beads come into proximity. The excitation of the Donor beads provokes the release of singlet oxygen molecules that triggers a cascade of energy transfer within the Acceptor beads, resulting in emission with λmax at 615 nm. The intensity of the signal is directly proportional to the concentration of analyte present in the sample.

Assay Principle AlphaLISA

 

Protocol of the AlphaLISA assay

The AlphaLISA assay can be run in a 96- or 384-well detection plate (50 µL final). As described here, samples or standards are dispensed directly into the assay plate for the detection of the analyte of interest by AlphaLISA reagents. No washing steps are needed. The protocol can be further miniaturized or upscaled by simply resizing each addition volume proportionally.

AlphaLISA 3-step assay protocol, acceptor bead first

Specifications

Application
Protein Quantification
Automation Compatible
Yes
Brand
AlphaLISA
Detection Modality
Alpha
Dynamic Range
5 - 100,000 pg/mL  
Limit of Detection
5 pg/mL  
Limit of Quantification
16 pg/mL  
Product Group
Kit
Sample Volume
10 µL
Shipping Conditions
Shipped in Blue Ice
Target
IL12
Target Class
Cytokines
Target Species
Porcine
Technology
Alpha
Therapeutic Area
Inflammation
Unit Size
100 assay points

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Application Note
Antigen-stimulated PBMC cytokine release measured by AlphaLISA bovine cytokine kits

Leptospirosis is an infectious disease caused by Leptospira bacteria, which can spread across different species of mammals...

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