Description
The d2 labeling kit is suitable for quick, easy labeling of peptides, proteins, and oligonucleotides with d2.
This kit is designed for very small-volume fast labeling needs, such as determining the best antibody among several antibodies available for HTRF assay development.
The N-hydroxysuccinimide activated d2 supplied reacts with primary amines carried by the biomolecules to label, for simplified conjugation.
The kit includes all the materials necessary for the labeling procedure (except dialysis and concentration devices).
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
The labeling procedure is performed under very mild conditions. As shown here, the process comprises an incubation step with the desiccated dye, followed by purification on a disposable column. Both the purification column and the buffer for purifying the conjugate are included in the kit, as well as a calibrator to define the ideal working dilution of the labeled material in HTRF assays. Depending on the molecular weight of the target molecule and the nature of the dye, each kit enables the labeling of 5-25 µg (protein-MW 15 kDa) to 50-250 µg (antibody-MW 150 kDa).
Assay protocol
The labeling procedure is described on the right. 100 µL of the targeted molecule (16.67 µM) in carbonate buffer pH 9.0 are added to the d2 vial. After an incubation of 45 minutes at RT, labeled protein is eluted on an equilibrated column. After having discarded 700 µL of elution buffer, the labeled molecule is collected in 400µL fraction.
