Revvity, 63ADK089PEG, HTRF Human Hyaluronic acid Detection Kit, 500 Assay Points

Hyaluronic acid (HA), also called hyaluronan, is an important component of the extracellular matrix contributing significantly to cell proliferation and migration. HA may also be involved in the progression of some malignant tumors. HA acts as a promoter of early inflammation, which is crucial to the whole skin wound-healing process. The hyaluronic acid kit is designed for a rapid detection of HA in cell supernatants. 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 Hyaluronic Acid is measured using a sandwich immunoassay involving Hyaluronic Acid Binding Protein (HABP) labelled with terbium Cryptate (donor) and HABP labelled with d2 (acceptor). The intensity of the signal is proportional to the concentration of Hyaluronic Acid present in the sample. Assay Protocol Cells are plated and stimulated in the same 96-well culture plate. Cell supernatants are then transferred into the assay plate for the detection of Hyaluronic Acid by HTRF reagents. This protocol enables the cells' viability and confluence to be monitored. The binding buffer is required to enhance the detection of Hyaluronic Acid on samples. The HABP labelled with HTRF fluorophores may be pre-mixed and added in a single dispensing step to further streamline the assay procedure. The assay detection can be run in 96- to 384-well plates by simply resizing each addition volume proportionally.