Revvity, AL3208C, AlphaLISA Human PICP (Pro-Collagen 1 C-Terminal Pro-Peptide) Detection Kit, 500 Assay Points

Overview
Collagen Type 1 is a protein present in abundance in the extracellular matrix, where forms cross-linking fibrils that associate in larger collagen fibers to play a role of structural support in connective tissues such as cartilage, bone, tendon, and skin. Due to its very unsoluble nature in its mature form, collagen is expressed and transported as a pro-collagen, in which the mature collagen sequence is flanked by two propetides respectively designated as PINP (Pro-collagen type 1 N-terminal Propeptide) and PICP (Pro-collagen type 1 C-terminal Propeptide). Mature collagen detection and quantification is not possible in immunoassays due to it being unsoluble and sticky. Instead, detection of the whole pro-collagen form or one of the propeptides is a good proxy to quantify the collagen expression in cells. In particular, targeting a propeptide like PICP allows to quantify both pro-collagen that have been expressed but not yet processed into mature collagen, and free PICP which have been cleaved from the mature sequence and are markers of collagen deposited in the extracellular matrix. The present kit is designed for the quantitative detection of Human PICP (Pro-Collagen 1 C-Terminal Pro-Peptide) in supernatants
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
Broad sample compatibility
Small sample volume
Results in less than 3 hours

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 human PICP assay
The AlphaLISA Human PICP assay is based on an AlphaLISA sandwich immunoassay involving a biotinylated anti-human PICP (hPICP) antibody bound to Streptavidin-coated AlphaLISA Donor beads and an anti-hPICP antibody conjugated to AlphaLISA Acceptor beads. Two antibodies are directed against the hPICP protein. In the presence of the hPICP, both antibodies bind to Human PICP 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 hPICP present in the sample (recombinant protein or cell supernatant).

Protocol of the AlphaLISA human PICP assay
The AlphaLISA Human PICP assay can be run in a 96 - or 384-well detection plate (50 µL final). As described here, samples (cell supernatants) or standards are dispensed directly into the assay plate for the detection of human PICP by AlphaLISA reagents. No washing steps are needed. The protocol can be further miniaturized or upscaled by simply resizing each addition volume proportionally.
Application-Protein Quantification
Automation Compatible-Yes
Brand-AlphaLISA
Detection Modality-Alpha
Dynamic Range-21 - 100,000 pg/mL
Limit of Detection-21 pg/mL
Limit of Quantification-66 pg/mL
Product Group-Kit
Protocol Time-1h30 at RT
Sample Volume-5 µL
Shipping Conditions-Shipped in Blue Ice
Target-PICP
Target Class-Biomarker
Target Species-Human
Technology-Alpha
Therapeutic Area-Oncology
Unit Size-500 assay points