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Human CTGF Kit HTRF®

The HTRF human CTGF kit is designed for the accurate measurement of human CTGF released into cell supernatants.

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  • No-wash No-wash
  • Ease-of-use Ease-of-use
  • Faster and more convenient than ELISA Faster and more convenient than ELISA
  • Highly accurate Highly accurate

The HTRF human CTGF kit is designed for the accurate measurement of human CTGF released into cell supernatants.

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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.

Benefits

  • Validated on human MRC5 and IMR90 cell lines.

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.

Principle of the HTRF CTGF assay

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

Assay protocol of the HTRF human CTGF

Technical specifications of human CTGF Kit

Sample size

16 µL

Final assay volume

20 µL

Kit components

Frozen standard and detection antibodies, buffers, & protocol.

LOD & LOQ (in Diluent)

 2 ng / mL & 4.2 ng / mL

Tidy

4.2 - 475 ng / mL

Time to result

Overnight at RT 

Calibration

No

Species

Human only

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.

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.

Validation of Human CTGF kit on human MRC5 cell lines

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Plate Reader Requirement

Choosing the right microplate reader ensures you’ll get an optimal readout. Discover our high performance reader, or verify if your lab equipment is going to be compatible with this detection technology.

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