Description
Human pro-collagen type 1 is made by a combined association of two pro-alpha1 chains with a pro-alpha2 chain. These triple-stranded, rope-like pro-collagen molecules must be processed by enzymes outside the cell to form fibrils that cross-link to produce strong mature type I collagen fibers. The human pro-collagen type 1 kit is designed for the quantitative detection of human pro-collagen type 1, using cell supernatants, whole cells, or recombinant protein extracts.
HTRF assays offer many advantages over other technologies:
Homogeneous add-and-read format
No wash steps
Low background
Straightforward miniaturization from 96- or 384-well microplates to high density assay formats such as 384-well low volume and 1536-well plates
Stable signal, providing flexibility in time of readout or size of assays
Assay principle
Human pro-collagen is measured using a sandwich immunoassay involving two specific anti-human pro-collagen antibodies, respectively labelled with Europium Cryptate (donor) and d2 (acceptor). The intensity of the signal is proportional to the concentration of pro-collagen present in the sample.
Assay protocol
The human pro-collagen type 1 assay protocol is shown here using a 384-well small volume white plate (20 µL final). Cell supernatant, sample, or standard is dispensed directly into the assay plate for the detection of human pro-collagen by HTRF reagents. The antibodies labelled with HTRF fluorophores may be pre-mixed and added in a single dispensing step to further streamline the assay procedure. The assay can be run in 96- to 384-well plates by simply resizing each addition volume proportionally.
