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AlphaLISA Human CTLA4 Detection Kit, 100 Assay Points

This product has been discontinued

The AlphaLISA™ immunoassay kit for human cytotoxic T-lymphocyte-associated protein 4 (CTLA-4) enables the quantitative determination of human CTLA-4 in buffered solution and cell culture supernatants using a homogeneous AlphaLISA assay (no wash steps).

Feature Specification
Application Protein Quantification
Dynamic Range 34 - 100,000 pg/mL  
Limit of Detection 34 pg/mL  
Limit of Quantification 121 pg/mL  
Sample Volume 10 µL

The AlphaLISA™ immunoassay kit for human cytotoxic T-lymphocyte-associated protein 4 (CTLA-4) enables the quantitative determination of human CTLA-4 in buffered solution and cell culture supernatants using a homogeneous AlphaLISA assay (no wash steps).

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

AlphaLISA technology allows the detection of molecules of interest in buffer, cell culture media, serum and plasma in a highly sensitive, quantitative, reproducible and user-friendly mode. In an AlphaLISA assay, a Biotinylated Anti-Analyte Antibody binds to the Streptavidin-coated Alpha 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 provokes 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.

Cytotoxic T-lymphocyte-associated protein 4 (CTLA4 or CTLA-4) is known as CD152 (cluster of differentiation 152). CTLA-4, as a protein receptor, is a critical biomarker for immune checkpoints. It is constitutively expressed in regulatory T cells (such as suppressor T cells) and the expression is upregulated in conventional T cells after activation. CTLA-4 acts as an "off" switch when bound to CD80 or CD86 on the surface of antigen-presenting cells. Thus, CTLA-4 is associated with numerous disease conditions such as insulin-dependent diabetes mellitus, systemic lupus erythematosus and other autoimmune diseases and cancers. Soluble CTLA-4 levels in biofluids have become a biomarker for determining the health status in patients. This kit is designed to detect and quantify the levels of CTLA-4 in cell culture supernatant, serum, and cell lysates.

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
34 - 100,000 pg/mL  
Limit of Detection
34 pg/mL  
Limit of Quantification
121 pg/mL  
Product Group
Kit
Sample Volume
10 µL
Shipping Conditions
Shipped in Blue Ice
Target
CTLA4
Target Class
Biomarkers
Target Species
Human
Technology
Alpha
Therapeutic Area
Oncology
Unit Size
100 assay points

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