Revvity, 64CTGFPEH, HTRF Human CTGF Detection Kit, 10,000 Assay Points

Connective tissue growth factor CTGF, also known as CCN2, is a matricellular signaling modulator. CTGF regulates a range of cellular functions, including cell adhesion, aggregation, and migration. CTGF modulates the balance of tissue remodeling in a large variety of cell types, such as vascular endothelial cells, fibroblasts, epithelial cells, and smooth muscle. CTGF is primarily regulated at the level of transcription and is induced by mechanical stresses and a number of cytokines and growth factors, including TGFβ. The HTRF human CTGF kit is a homogeneous add and read assay, to measure CTGF secretion in cell supernatants.
CTGF Assay Principle CTGF is measured using a sandwich immunoassay involving two specific anti-CTGF antibodies, respectively labeled with HTRF donor and acceptor dyes. The intensity of the signal is directly proportional to the concentration of CTGF present in the sample. CTGF Assay Protocol Cell supernatant, sample, or standard is dispensed directly into the assay plate for CTGF detection by HTRF reagents (384-well low-volume white plate in 20 µL). The antibodies labeled with the HTRF donor and acceptor are pre-mixed and added in a single dispensing step, to further streamline the assay procedure
CTGF secretion in MRC5 cell line stimulated with TGFbeta Human MRC5 cells were plated at 400,000 cells/well in a 12-well plate and cultured overnight with 600 µL of complete MEM cell culture medium, at 37°C, 5% CO2. Cells were then stimulated with increasing concentrations of TGFbeta (0.2 to 20 ng/mL) under 600 µL of MEM + 1%. After 24h of stimulation, 16 µL of supernatant were transferred into a white detection plate to be analyzed using the Human CTGF assay. The HTRF signal was recorded after an overnight incubation.