Revvity, 62HPIL1BPEG, HTRF Human pro IL-1β Detection Kit, 500 assay points

Overview
Pro IL-1β, also known as IL-1β precursor, is a 31 kDa protein precursor of IL-1β. It is produced in response to molecular motifs carried by pathogens called ‘pathogen associated molecular patterns’ (PAMPs). Induction of pro IL-1β expression is referred to as a priming step and is an inefficient secretion stimulus. The processing and cleavage of pro IL-1β into active IL-1β is conditioned by the activation stage involving PAMP, or DAMP (danger associated molecular pattern, endogenous molecules released from dead cells).
Pro IL-1β is cleaved by the pro-inflammatory protease caspase-1, activated via recruitment to a multi-protein complex termed the inflammasome.
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 readout time or size of assays
How it works
Principle of the HTRF human pro IL-1β assay
The HTRF pro IL-1β assay is based on a TR-FRET sandwich immunoassay involving two specific antibodies, one labeled with Eu3+-cryptate (donor) and the other with d2 (acceptor). One antibody is directed against the IL-1β, and the second recognizes specifically pro IL-1β. When both antibodies bind to pro IL-1β, and the donor-acceptor proximity enables a fluorescent TR-FRET signal. The intensity of the signal is directly proportional to the concentration of pro IL-1β present in the cell lysate sample.

 

Protocol of the HTRF human pro IL-1β assay
The HTRF pro IL-1β assay can be run in a 96- or 384-well low volume white detection plate (20 µL final). As described here, samples (cell lysates) or standards are dispensed directly into the assay plate for the detection of pro IL-1β by HTRF reagents. The antibodies labelled with HTRF fluorophores may be pre-mixed and added in a single dispensing step. No washing steps are needed. The protocol can be further miniaturized or upscaled by simply resizing each addition volume proportionally. Application-Protein Quantification
Brand-HTRF
Detection Modality-HTRF
Dynamic Range-312.5 - 20,000 pg/mL
Limit of Detection-38.5 pg/mL
Limit of Quantification-130 pg/mL
Product Group-Kit
Sample Volume-16 µL
Shipping Conditions-Shipped in Dry Ice
Target-pro-IL1β
Target Class-Cytokines
Target Species-Human
Technology-TR-FRET
Therapeutic Area-Autoimmunity Cardiovascular Immuno-oncology Infectious Diseases Inflammation Neuroscience Oncology Virology
Unit Size-500 assay points