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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
3.0 hours or 4.5 hours
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Sample Type & Volume Required Per Well
Cell Culture Supernates (10 uL), Serum (150 uL), EDTA Plasma (150 uL), Heparin Plasma (150 uL), Citrate Plasma (150 uL)
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Sensitivity
11 pg/mL
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Assay Range
15.6 - 1,000 pg/mL (Cell Culture Supernates), 31.2 - 2,000 pg/mL (Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma)
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Specificity
Natural and recombinant human MIP-1 beta
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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
QC20, Quantikine Immunoassay Control Group 3 -
Product Summary
The Quantikine Human MIP-1 beta Immunoassay is a 3.0 or 4.5 hour solid phase ELISA designed to measure MIP-1 beta in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant human MIP-1 beta (Act-2 variant) and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant human MIP-1 beta. Results obtained using natural human MIP-1 beta 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 MIP-1 beta.
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.
Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma | Intra-Assay Precision | Inter-Assay Precision |
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 40 | 40 | 40 |
| Mean | 51.3 | 208 | 932 | 58.8 | 220 | 949 |
| Standard Deviation | 4.6 | 7.5 | 30.2 | 5.7 | 15.6 | 70.5 |
| CV% | 9 | 3.6 | 3.2 | 9.7 | 7.1 | 7.4 |
Cell Culture Supernates | Intra-Assay Precision | Inter-Assay Precision |
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 40 | 40 | 40 |
| Mean | 23.3 | 98.8 | 428 | 31.4 | 118 | 476 |
| Standard Deviation | 1.1 | 3.8 | 17.3 | 3.7 | 9.9 | 28 |
| CV% | 4.7 | 3.8 | 4 | 11.8 | 8.4 | 5.9 |
Recovery
The recovery of MIP-1 beta spiked to three levels in samples throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
| Cell Culture Media (n=5) | 107 | 81-117 |
| Citrate Plasma (n=5) | 99 | 87-116 |
| EDTA Plasma (n=5) | 100 | 84-112 |
| Heparin Plasma (n=5) | 99 | 90-112 |
| Serum (n=5) | 99 | 88-113 |
Linearity
To assess the linearity of the assay, five samples were spiked with high concentrations of MIP-1 beta in various matrices and diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Preparation and Storage
Background: CCL4/MIP-1 beta
CCL4/MIP-1 beta is a beta chemokine that is secreted at sites of inflammation by activated leukocytes, lymphocytes, vascular endothelial cells, and pulmonary smooth muscle cells. It attracts a variety of immune cells to sites of microbial infection as well as to other pathologic inflammation such as allergic asthma and ischemic myocardium. CCL4 is secreted from activated monocytes as a heterodimer with CCL3/MIP-1 alpha. It signals through CCR5, and an N-terminally trimmed form additionally interacts with CCR1 and CCR2. In humans, the ability of CCL4 to bind CCR5 inhibits the cellular entry of M-tropic HIV-1 which utilizes CCR5 as a coreceptor.
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