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HTRF Human CTGF Detection Kit, 500 Assay Points
HTRF Human CTGF Detection Kit, 500 Assay Points
Shipping box for Revvity reagent kits
The HTRF human CTGF kit is designed for the accurate measurement of human CTGF released into cell supernatants.
| Feature | Specification |
|---|---|
| Application | Protein Quantification |
| Sample Volume | 16 µL |
The HTRF human CTGF kit is designed for the accurate measurement of human CTGF released into cell supernatants.
Product Variants
Unit Size: 500 Assay Points
Part #:
64CTGFPEG
List Price
USD 1,671.37
Unit Size: 10,000 Assay Points
Part #:
64CTGFPEH
List Price
USD 19,683.38
For research use only. Not for use in diagnostic procedures. All products to be used in accordance with applicable laws and regulations including without limitation, consumption and disposal requirements under European REACH regulations (EC 1907/2006).
HTRF Human CTGF Detection Kit, 500 Assay Points
Shipping box for Revvity reagent kits
HTRF Human CTGF Detection Kit, 500 Assay Points
Product information
Overview
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.
Specifications
| Application |
Protein Quantification
|
|---|---|
| Brand |
HTRF
|
| Detection Modality |
HTRF
|
| Product Group |
Kit
|
| Sample Volume |
16 µL
|
| Shipping Conditions |
Shipped in Dry Ice
|
| Target Class |
Biomarkers
|
| Target Species |
Human
|
| Technology |
TR-FRET
|
| Therapeutic Area |
NASH/Fibrosis
|
| Unit Size |
500 Assay Points
|
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HTRF Human CTGF Detection Kit, 500 Assay Points
HTRF Human CTGF Detection Kit, 500 Assay Points
How it works
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
Analytical performance
Intra / Inter assay
Intra-assay (n=24)
| Sample | Mean [CTGF] (ng/mL) | CV |
|---|---|---|
| 1 | 10.1 | 14.7 |
| 2 | 69.25 | 10.6 |
| 3 | 475 | 10 |
| Mean CV | 11.7 |
Each of the 3 samples was measured 24 times, and the %CV for the concentration was calculated for each sample
Inter-assay (n=4)
| Sample | [CTGF] (ng/mL) | Mean (delta R) | CV |
|---|---|---|---|
| 1 | 19.2 | 1100 | 5 |
| 2 | 131.6 | 14922 | 11 |
| 3 | 250 | 24765 | 2 |
| Mean CV | 6 |
Each of the samples was measured in 4 different experiments, and the %CV for the concentration was calculated for each sample.
Spike and recovery
| Sample | [CTGF] recomb prot (ng/mL) | [CTGF] native prot (ng/mL) | [CTGF]expected (pg/mL) | [CTGF]detected (pg/mL) | Recovery | ||
|---|---|---|---|---|---|---|---|
| 1 | 14.02 | 15.02 | 29.04 | 34.73 | 120 % | ||
| 2 | 14.02 | 22.05 | 36.07 | 39.1 | 108% | ||
| 3 | 25.19 | 16.88 | 42.07 | 42.07 | 110% |
Different amounts of recombinant CTGF were added to MEM + 1% BSA cell supernatant samples containing a known concentration of CTGF . Samples were mixed at a 1:1 volume ratio. The set of responses obtained from a standard curve was compared to the calculated expected values. The recovery value observed validates the sample matrix used for this assay.
Assay validation
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.
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