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AlphaLISA Human VEGF Detection Kit, 500 Assay Points

The AlphaLISA™ immunoassay kit for detection and quantitation of human VEGF (VEGF-A) in serum, cell culture supernatants, and other sample types allows for fast, reproducible, and sensitive detection without the need for time-consuming wash steps or complicated assay development.

Feature Specification
Application Protein Quantification
Dynamic Range 2.2 - 100,000 pg/mL
Limit of Detection 2.2 pg/mL  
Sample Volume 5 µL

The AlphaLISA™ immunoassay kit for detection and quantitation of human VEGF (VEGF-A) in serum, cell culture supernatants, and other sample types allows for fast, reproducible, and sensitive detection without the need for time-consuming wash steps or complicated assay development.

Product variants
Unit Size: 500 assay points
Part #:
AL201C
List price
USD 4,722.55
Your online price:
Unit Size: 5,000 assay points
Part #:
AL201F
List price
USD 36,853.40
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

The antibodies in this kit recognize VEGF-A (VEGF165 or isoform d), VEGF162, VEGF145 (isoform e) and VEGF121 (isoform f). One of the sandwiching antibodies is a mouse monoclonal, clone 26503. The other sandwiching antibody is a goat polyclonal raised against human recombinant VEGF165.

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

Vascular endothelial growth factor (VEGF-A or VEGF) is a homodimeric 34-42 kDa heparin-binding glycoprotein specific for endothelial cells. Currently at least eight variants of VEGF generated by alternative splicing of the same gene have been identified. The various isoforms have different heparin- and neuropilin(-1 or -2)-binding properties as well as solubility characteristics reflecting the different functional properties of the VEGF forms. VEGF is believed to play important roles in inflammation and during normal and pathological angiogenesis, a process that is associated with wound healing, embryonic development, and growth and metastasis of solid tumors. Elevated levels of VEGF have been observed in synovial fluids of rheumatoid arthritis patients and sera of cancer patients.

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
2.2 - 100,000 pg/mL
Limit of Detection
2.2 pg/mL  
Product Group
Kit
Sample Volume
5 µL
Shipping Conditions
Shipped in Blue Ice
Target
VEGF-A
Target Class
Biomarkers
Target Species
Human
Technology
Alpha
Therapeutic Area
Oncology
Unit Size
500 assay points

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Application Note
Alpha technology: a fast and sensitive orthogonal approach to cell-based potency assays

Orthogonal systems to cell-based assays are a key requirement in EMA/FDA guidelines for potency estimations and require cross...

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