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  • AlphaLISA HIV p24 Biotin Free Detection Kit, 100 Assay Points

AlphaLISA HIV p24 Biotin Free Detection Kit, 100 Assay Points

AlphaLISA Biotin Free image
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AlphaLISA HIV p24 Biotin Free Detection Kit, 100 Assay Points
AlphaLISA Biotin Free image
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AlphaLISA Biotin Free image
AlphaLISA Biotin Free
AlphaLISA Biotin Free image
AlphaLISA Biotin Free image
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The AlphaLISA HIV p24 Biotin-Free Detection Kit is designed for the quantitative detection of HIV p24 in serum, cell culture medium, and other samples types using a homogeneous (no wash steps, no separation steps) assay. The biotin-free kit uses anti-DIG (anti-Digoxin) Donor beads instead of streptavidin Donor beads, which makes the kit compatible with high-biotin culture media and other sample types that contain high levels biotin (including brain/liver tissue extracts, milk and eggs).

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Feature Specification
Application Protein Quantification
Dynamic Range 10 - 100,000 pg/mL  
Limit of Detection 9.6 pg/mL  
Limit of Quantification 35.4 pg/mL  
Sample Volume 10 µL

The AlphaLISA HIV p24 Biotin-Free Detection Kit is designed for the quantitative detection of HIV p24 in serum, cell culture medium, and other samples types using a homogeneous (no wash steps, no separation steps) assay. The biotin-free kit uses anti-DIG (anti-Digoxin) Donor beads instead of streptavidin Donor beads, which makes the kit compatible with high-biotin culture media and other sample types that contain high levels biotin (including brain/liver tissue extracts, milk and eggs).

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Product variants
Unit Size: 100 assay points
Part #:
AL332HV
Unit Size: 500 assay points
Part #:
AL332C
Unit Size: 5,000 assay points
Part #:
AL332F
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).
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AlphaLISA Biotin Free image
AlphaLISA Biotin Free image
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AlphaLISA HIV p24 Biotin Free Detection Kit, 100 Assay Points
AlphaLISA Biotin Free image
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AlphaLISA Biotin Free image
AlphaLISA Biotin Free
AlphaLISA HIV p24 Biotin Free Detection Kit, 100 Assay Points
AlphaLISA Biotin Free image
AlphaLISA Biotin Free image
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Product information

  • Overview
  • How it works
  • Specifications
  • Video gallery

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 and Benefits:

  • No-wash steps, no separation steps
  • Compatible with high biotin samples, including RPMI and high-biotin tissue extracts
  • ELISA alternative technology
  • Sensitive detection
  • Broad sample compatibility
  • Results in less than 3 hours

Immunodeficiency Virus (HIV) p24 is the 231 amino acid phosphorylated protein of the capsid forming the conical core of the virus that encapsulates the genomic RNA-nucleocapsid complex. HIV p24 is a cleavage product of the p55 Gag polyprotein by viral proteases. HIV p24 and its 55 kDa precursor play a crucial role in the assembly, maturation, and disassembly of HIV p24 can often be detected two weeks after infection. Subsequently, p24 antibody is produced and complexes with soluble p24 antigen, rendering it undetectable without first dissociating the antibody-antigen complex. Free antigen reappears later in the course of the illness as p24 antibody levels decline. p24 is frequently used for HIV detection in blood, serum samples, and other bodily fluids in acute HIV seroconversion, in neonatal infection, and for monitoring of responses to antiviral drug therapy.

AlphaLISA technology allows the detection of molecules of interest in a no-wash, highly sensitive, quantitative assay. In an AlphaLISA biotin-free assay, a DIG-labeled anti-analyte antibody binds to the anti-DIG-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 biotin-free assay

The AlphaLISA Biotin-Free assay is based on an AlphaLISA sandwich immunoassay involving a digoxigenin anti-analyte antibody bound to anti-DIG 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.

AlphaLISA DIG Assay Principle

 

Protocol of the AlphaLISA biotin-free assay

The AlphaLISA Biotin-Free 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.

img-protocol-alphalisa-2-step-dig

Specifications

Application
Protein Quantification
Automation Compatible
Yes
Brand
AlphaLISA
Detection Modality
Alpha
Dynamic Range
10 - 100,000 pg/mL  
Limit of Detection
9.6 pg/mL  
Limit of Quantification
35.4 pg/mL  
Product Group
Kit
Sample Volume
10 µL
Shipping Conditions
Shipped in Blue Ice
Target
p24
Target Class
Biologics
Technology
Alpha
Therapeutic Area
Virology
Unit Size
100 assay points

Video gallery

AlphaLISA HIV p24 Biotin Free Detection Kit, 100 Assay Points
AlphaLISA HIV p24 Biotin Free Detection Kit, 100 Assay Points

Resources

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Application Note
Avoiding biotin interference in AlphaLISA assays

AlphaLISA™ technology is a highly sensitive, easy-to-use, and reproducible method for detecting and quantifying molecules in...

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