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Mouse CCL2/JE/MCP-1 Quantikine ELISA Kit

R&D Systems, part of Bio-Techne | Catalog # MJE00

R&D Systems, part of Bio-Techne
Discontinued Product
MJE00 has been discontinued. An alternative/replacement product is available: MJE00B. View all CCL2/JE/MCP-1 products.

Key Product Details

Assay Length

4.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (25 µL), Serum (25 µL)

Sensitivity

2 pg/mL

Assay Range

15.6-1000 pg/mL (Cell Culture Supernates, Serum)

Product Summary for Mouse CCL2/JE/MCP-1 Quantikine ELISA Kit

The Quantikine Mouse/Rat CCL2/MCP-1 Immunoassay is a 4.5 hour solid phase ELISA designed to measure MCP-1 in mouse or rat cell culture supernates and serum. It contains E. coli-expressed mouse MCP-1 and antibodies raised against the recombinant factor. This Immunoassay has been shown to accurately quantitate recombinant mouse MCP-1. Results obtained using natural mouse or rat MCP-1 showed dose response curves that were parallel to the standard curves obtained using the Quantikine mouse kit standards. These results indicate that this kit can be used to determine relative mass values for natural MCP-1.

Product Specifications

Assay Type

Solid Phase Sandwich ELISA

Format

96-well strip plate

Measurement

Quantitative ELISA

Detection Method

Colorimetric - 450nm (TMB)

Conjugate

HRP

Reactivity

Mouse, Rat

Specificity

Natural and recombinant mouse and rat MCP-1

Cross-reactivity

< 0.5% cross-reactivity observed with available related molecules. < 50% cross-species reactivity observed with species tested.

Interference

No significant interference observed with available related molecules.

Sample Values

Serum - Mouse and rat samples were evaluated for the presence of MCP-1 in this assay.


Mean (pg/mL)Range (pg/mL)Standard Deviation (pg/mL)
Mouse serum* (n=40)13050-29447
Rat serum* (n=10)1346768-1584278
*Samples were diluted prior to assay.

Cell Culture Supernates:
Mouse heart tissue was cut into 1-2 mm pieces and cultured in 10 mL of RPMI supplemented with 10% fetal bovine serum for 7 days. An aliquot of the cell culture supernate was removed, assayed for levels of mouse MCP-1, and measured 3000 pg/mL.

L-929 mouse fibroblasts (1 x 105 cells/mL) were cultured for 3 days in MEM containing L-glutamine and 10% equine serum. An aliquot of the cell culture supernate was removed, assayed for levels of mouse MCP-1 and measured 13,000 pg/mL.

Rat splenocytes were cultured for 22 hours in RPMI supplemented with 10% fetal bovine serum. The cells were stimulated with 10 μg/mL of PHA and 10 ng/mL of PMA. An aliquot of the cell culture supernate was removed, assayed for levels of rat MCP-1 and measured 891 pg/mL.

Precision

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.

Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.

Cell Culture Supernates, Serum

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (pg/mL) 47.1 311 768 43.6 310 783
Standard Deviation 3.9 15.8 40.6 3.2 14.3 42.8
CV% 8.3 5.1 5.3 7.3 4.6 5.5

Recovery for Mouse CCL2/JE/MCP-1 Quantikine ELISA Kit

The recovery of MCP-1 spiked to three levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Mouse Cell Culture Supernates (n=6) 104 91-119
Mouse Serum (n=9) 99 89-113
Rat Cell Culture Supernates (n=4) 109 103-115
Rat Serum (n=4) 109 103-116

Linearity

To assess the linearity of the assay, samples containing and/or spiked with high concentrations of MCP-1 in each matrix were diluted with Calibrator Diluent and then assayed.

Mouse/Rat CCL2/JE/MCP-1 ELISA Linearity
Mouse CCL2/JE/MCP-1 ELISA Linearity

Scientific Data Images for Mouse CCL2/JE/MCP-1 Quantikine ELISA Kit

Mouse/Rat CCL2/JE/MCP-1 Standard Curve

Mouse/Rat CCL2/JE/MCP-1 Standard Curve

Mouse CCL2/JE/MCP-1 ELISA Standard Curve

Mouse CCL2/JE/MCP-1 ELISA Standard Curve

Preparation and Storage

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: CCL2/JE/MCP-1

CCL2/JE/MCP-1, is a chemokine that binds the receptor CCR2 and induces the chemoattraction of mononuclear cells. It induces the activation of monocytes, NK cells, lymphocytes, and basophils. Additionally, CCL2 promotes Th2 polarization in CD4+ T cells, and CCL2-mediated recruitment of monocytes to sites of inflammation contributes to disease severity in atherosclerosis, multiple sclerosis, and allergic asthma. Endogenous proteolytic trimming of CCL2 at the N-terminus, including the N-terminal pyrrolidone carboxylic acid-modified glutamine, downregulates activity but not receptor binding.

Alternate Names

GDCF-2, HC11, HSMCR30, MCAF, MCP-1, SMC-CF

Entrez Gene IDs

6347 (Human); 20296 (Mouse); 24770 (Rat); 403981 (Canine)

Gene Symbol

CCL2

Additional CCL2/JE/MCP-1 Products

Product Documents for Mouse CCL2/JE/MCP-1 Quantikine ELISA Kit

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Mouse CCL2/JE/MCP-1 Quantikine ELISA Kit

For research use only

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⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.