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.
Mouse CCL2/JE/MCP-1 Quantikine ELISA Kit
R&D Systems | Catalog # MJE00
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)
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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
Species
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.
*Samples were diluted prior to assay.
| Mean (pg/mL) | Range (pg/mL) | Standard Deviation (pg/mL) | |
| Mouse serum* (n=40) | 130 | 50-294 | 47 |
| Rat serum* (n=10) | 1346 | 768-1584 | 278 |
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.
Scientific Data Images for Mouse CCL2/JE/MCP-1 Quantikine ELISA Kit
Mouse/Rat CCL2/JE/MCP-1 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
Alternate Names
GDCF-2, HC11, HSMCR30, MCAF, MCP-1, SMC-CF
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 or batch 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
⚠ 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.Citations for Mouse CCL2/JE/MCP-1 Quantikine ELISA Kit
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Sample Tested: Mouse lung tissueVerified Customer | Posted 05/19/2017This ELISA worked perfectly in the lung lysates of Scistosoma induced PH mouse model.Thanks
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Protocols
View specific protocols for Mouse CCL2/JE/MCP-1 Quantikine ELISA Kit (MJE00):
Refer to the product for complete assay procedure.
Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
- Prepare all reagents, standard dilutions, and samples as directed in the product insert.
- Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
- Add 50 µL of Assay Diluent to each well.
- Add 50 µL of Standard, Control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
- Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.
- Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
- Aspirate and wash 5 times.
- Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.
- Add 100 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
50 µL Assay Diluent
50 µL Standard, Control, or Sample
100 µL Conjugate
100 µL Substrate Solution
100 µL Stop Solution
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
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