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Assay Type
Solid Phase Sandwich ELISA
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Format
96-well strip plate
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Assay Length
3.5 hours or 4.5 hours
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Sample Type & Volume Required Per Well
Cell Culture Supernates (200 uL), Serum (100 uL), EDTA Plasma (100 uL), Heparin Plasma (100 uL), Citrate Plasma (100 uL), Urine (100 uL)
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Sensitivity
10 pg/mL
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Assay Range
31.2 - 2,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma, Urine)
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Specificity
Natural and recombinant human MCP-1
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Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
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Interference
No significant interference observed with available related molecules.
Control Available
QC01-1, Quantikine Immunoassay Control Group 1 -
Product Summary
The Quantikine Human MCP-1 Immunoassay is a 3.5 or 4.5 hour solid phase ELISA designed to measure MCP-1 in cell culture supernates, serum, plasma, and urine. It contains E. coli-expressed recombinant human MCP-1 and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate recombinant human MCP-1. Results obtained using natural human MCP-1 showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural human MCP-1.
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, Urine | Intra-Assay Precision | Inter-Assay Precision |
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean | 78.2 | 360 | 1079 | 76 | 360 | 1086 |
| Standard Deviation | 3.3 | 17.7 | 63.5 | 4.5 | 17.2 | 48.5 |
| CV% | 4.2 | 4.9 | 5.9 | 5.9 | 4.8 | 4.5 |
Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma | Intra-Assay Precision | Inter-Assay Precision |
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean | 76.7 | 364 | 1121 | 74.2 | 352 | 1076 |
| Standard Deviation | 6 | 17.1 | 54.4 | 5 | 20.5 | 49.4 |
| CV% | 7.8 | 4.7 | 4.9 | 6.7 | 5.8 | 4.6 |
Recovery
The recovery of MCP-1 spiked to three levels throughout the range of the assay was evaluated.
| Sample Type | Average % Recovery | Range % |
| Cell Culture Media (n=5) | 96 | 88-107 |
| Citrate Plasma (n=5) | 100 | 94-107 |
| EDTA Plasma (n=5) | 96 | 92-102 |
| Heparin Plasma (n=5) | 102 | 94-114 |
| Serum (n=5) | 103 | 92-113 |
| Urine (n=5) | 92 | 85-100 |
Linearity
To assess the linearity of the assay, samples were spiked with high concentrations of MCP-1 and diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Preparation and Storage
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.
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Alternate Names
CCL2; MCAF; MCP-1;
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