Mouse CCL2 (MCP1) kit HTRF®

Name: Mouse CCL2 (MCP1) kit
Brand: Cisbio
SKU: 40456
Availability: InStock

Associated products

The HTRF mouse CCL2 (MCP1) kit is designed for the quantification of mouse CCL2 release in cell supernatant.

No-wash
Low sample consumption
Under 2h bench time

Download the protocols

The HTRF mouse CCL2 (MCP1) kit is designed for the quantification of mouse CCL2 release in cell supernatant.

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Overview

Also called MCP-1 (monocyte chemoattractant protein 1), CCL2 is a chemokine mainly secreted by monocytes, macrophages, and dendritic cells to attract monocytes, basophils, and T cells. CCL2 is involved in pathologies like psoriasis or atherosclerosis. In the brain, CCL2 is released by neurons, astrocytes, and microglia, and is part of the neuro-inflammatory response observed in 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.

Assessment of serum samples often requires enhanced sensitivity. In some cases, AlphaLISA assays may have sufficient sensitivity to enable the detection of low levels of analytes in serum or plasma. Check out the Mouse/rat CCL2/MCP1 AlphaLISA kit’s analytical performances for more information or learn more about AlphaLISA. When assaying, always follow the recommended protocol and avoid highly haemolyzed samples.

Benefits

SCALABLE FROM 96 TO 1536WELLS
PANEL QUANTIFICATION

Assay principle

Cell supernatant, sample, or standard is dispensed directly into the assay plate for the detection by HTRF® reagents (384-well low-volume white plate or Cisbio low-volume 96-well 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. The assay can be run up to a 1536-well format by simply resizing each addition volume proportionally.

Assay data analysis

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.

To fully understand how to deal with HTRF data processing and also 4PL 1/y² fitting, please visit this page.

Cisbio also worked with Myassays.com to help you in your data analysis. By clicking on this link, you’ll be able to access a free online software to run your CCL2 analysis.

Technical specifications of mouse CCL2 (MCP1) kit

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

Intra and inter assay

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%

CCL2 secretion in 3T3-L1 cells stimulated with IL1ß and TNFa

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 IL1ß and TNFa, both used at 10ng/ml. Supernatants were diluted 100 times. 16 µL

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.

CCL2 secretion in 3T3-L1 cells stimulated with IL1ß and TNFa.

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