Mouse CCL2 (MCP1) Assay Kit

Also called MCP-1 (monocyte chemoattractant protein 1), CCL2 is a chemokine mainly secreted by monocytes, macrophages and dendritic cells. Its main role is to attract monocytes, basophils and T cells.

CCL2 is involved in pathologies where monocytic infiltrates are found, like psoriasis or atherosclerosis.

In the brain, CCL2 is released by neurons, astrocytes, and microglia, and is part of the neuroinflammatory response observed in epilepsy, ischemia or Alzheimer’s disease.

CCL2 is also secreted by adipocytes and can interfere with insulin signaling, thus suggesting a role in the inflammation linked to Diabetes.

Cisbio's Mouse CCL2 kit is a homogeneous, add and read assay to measure CCL2 secretion in supernatant. It offers a fast and efficient alternative to ELISA

Sample size 16 µL
Final assay volume 20 µL
Kit components Lyophilized standard, frozen detection antibodies, buffers & protocol
LOD & LOQ (in Diluent) 6 pg/mL & 13 pg/mL
Range 13 – 1,800 pg/mL
Time to result ON at RT
Calibration NA
Species Mouse only
Download Insider tips for PBMC isolation Technical Note
Download cytokine release from fresh blood samples application notes

Assay principle

The assay protocol, using a Cisbio 96-well low volume or a 384-well small volume white plate (20 µL final), is described below. Cell supernatant, sample or standard are dispensed directly into the assay plate for the detection by HTRF® reagents. 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. The assay can be run in 96 to 1536-well plates by simply resizing each addition volume proportionally.

The 4 Parameter Logistic (4PL) curve is commonly recommended for fitting an ELISA standard curve. This regression enables the accurate measurement of an unknown sample across a wider range of concentrations than linear analysis, making it ideally suited to the analysis of biological systems like cytokine releases.

Add

Cytokines assay principle add

Read

Cytokines assay principle read

(1) Assay data analysis:

  • "Guidelines for data reduction" button will show you how to deal with HTRF data processing and 4PL 1/y2 fitting.
  • "MyAssays" button will direct you to a free online software able to run IFNγ analysis.

 

How to use HTRF cytokine assays

How to measure multiple cytokines from 1 sample

Analytical performance

Precision

Intra assay (n=24)

Sample Mean [CCL2] (pg/mL) CV
1 173 5%
2 561 2%
3 1594 2%
  Mean CV 3%

Inter assay (n=4)

Sample [CCL2] (pg/ml) Mean (delta ratio) CV
1 72 722 10%
2 263 3102 11%
3 960 12960 16%
    Mean CV 12%

Assay validation

CCL2 secretion in 3T3-L1 cells stimulated with IL1β and TNFα.

3T3-L1 cells* were plated in a 12 well plate at 30,000 cells per well under 1ml. Cells were grown until confluency, and differentiated onto adipocyte (~700,000 cells per well). Differentiated cells were then stimulated for 24h with IL1β and TNFα, both used at 10ng/ml. Supernatants were diluted 100 times. 16µl of diluted supernatants were transferred into a 384 well (SV) white detection plate to be analyzed by the Mouse CCL2 Assay Kit.

*Samples were kindly provided by J.F. Tanti’s research team: Cellular and Molecular Physiopathology of Obesity, Inserm U1065, Nice, FRANCE.

Part#, inserts & MSDS

Ordering Info

DescriptionCat. noProduct insertMSDS
Mouse CCL2 kit - 500 tests62MCCL2PEG
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Mouse CCL2 kit - 10,000 tests62MCCL2PEH
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Companion products

DescriptionCat. noProduct insertMSDS
Mouse CCL2 kit - 3 Standards62MCCL2CD3
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Mouse CCL2 kit - Standard62MCCL2CDA
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