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AlphaLISA Human Factor VIII Detection Kit, 5,000 Assay Points

The AlphaLISA™ Human VIII Detection Kit is designed for detection and quantitation of human VIII in cell culture media or serum using a homogeneous (no-wash steps, no separation steps) assay.

Feature Specification
Application Protein Quantification
Dynamic Range 87 - 100,000 pg/mL  
Limit of Detection 87 pg/mL  
Limit of Quantification 284 pg/mL  
Sample Volume 5 µL

The AlphaLISA™ Human VIII Detection Kit is designed for detection and quantitation of human VIII in cell culture media or serum using a homogeneous (no-wash steps, no separation steps) assay.

Product variants
Unit Size: 100 assay points
Part #:
AL342HV
List price
USD 1,044.42
Your online price:
Unit Size: 500 assay points
Part #:
AL342C
List price
USD 2,533.13
Your online price:
Unit Size: 5,000 assay points
Part #:
AL342F
List price
USD 16,428.50
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 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:

  • 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

Factor VIII is a blood clotting protein which is also called anti-hemophilic factor (AHF). Factor VIII is produced in the liver and in the endothelial cells of other organs and tissues. It circulates in the bloodstream as an inactive form complexed with von Willebrand factor (vWF). In damaged blood vessels Factor VIII disassociates from vWF to become active Factor VIII. Activated Factor VIII initiates the cascade of chemical reactions to promote blood clotting at the site of vessel damage to prevent blood loss from the body. For this reason, it has been used as a blood clotting drug for patients with hemophilia to restore hemostasis. The present kit detects hFactor VIII in human serum, plasma, cell culture supernatants, and cell lysate and tissue homogenates.

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
87 - 100,000 pg/mL  
Limit of Detection
87 pg/mL  
Limit of Quantification
284 pg/mL  
Product Group
Kit
Sample Volume
5 µL
Shipping Conditions
Shipped in Blue Ice
Target
Factor VIII
Target Class
Biomarkers
Target Species
Human
Technology
Alpha
Therapeutic Area
Oncology
Unit Size
5,000 assay points

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