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Assay Length
3.5 hours
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Sample Type & Volume Required Per Well
Cell Culture Supernates (100 uL), Serum (100 uL), EDTA Plasma (100 uL), Heparin Plasma (100 uL), Citrate Plasma (100 uL), Urine (100 uL)
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Sensitivity
6.6 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 RANTES
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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 RANTES Immunoassay is a 3.5 hour solid phase ELISA designed to measure human RANTES levels in cell culture supernates, serum, plasma, and urine. It contains E. coli-derived recombinant human RANTES and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant factor. Results obtained using natural human RANTES 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 of natural human RANTES.
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, Urine | Intra-Assay Precision | Inter-Assay Precision |
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean | 91.9 | 573 | 1120 | 92 | 511 | 1105 |
| Standard Deviation | 2.3 | 10 | 32.4 | 6.2 | 32.7 | 70.7 |
| CV% | 2.5 | 1.7 | 2.9 | 6.7 | 6.4 | 6.4 |
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 | 108 | 599 | 1191 | 88 | 494 | 1063 |
| Standard Deviation | 3.9 | 14.3 | 41.5 | 9.1 | 43.1 | 89.6 |
| CV% | 3.6 | 2.4 | 3.5 | 10.3 | 8.7 | 8.4 |
Recovery
The recovery of RANTES spiked to levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
| Cell Culture Media (n=4) | 101 | 95-110 |
| Platelet-poor Citrate Plasma (n=4) | 99 | 93-105 |
| Platelet-poor EDTA Plasma (n=4) | 98 | 93-103 |
| Platelet-poor Heparin Plasma (n=4) | 99 | 93-105 |
| Serum (n=5) | 101 | 94-107 |
| Urine (n=10) | 102 | 88-114 |
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of RANTES were serially diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Preparation and Storage
Background: CCL5/RANTES
CCL5/RANTES is a secreted beta-chemokine that plays a primary role in the inflammatory immune response through chemoattraction and activation of leukocytes that express CCR1, 3, 4, or 5. The two N-terminal residues of CCL5 can be removed by CD26/DPPIV, generating a protein that functions as a chemotaxis inhibitor. The deposition of CCL5 on activated vascular endothelial cells is crucial for monocyte adhesion to damaged vasculature. CCL5 is upregulated in breast cancer and promotes tumor progression through the attraction of proinflammatory macrophages in addition to its actions on tumor cells, stromal cells, and the vasculature. In humans, CCR5 binding to CCL5 inhibits the infectivity of R5 (M-tropic) but not X4 (T-tropic) strains of HIV-1.
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Alternate Names
CCL5; chemokine (C-C motif) ligand 5; D17S136Enormally T-expressed, and presumably secreted; EoCP; Eosinophil chemotactic cytokine; RANTES; SISd; SIS-delta; small inducible cytokine A5 (RANTES); small inducible cytokine subfamily A (Cys-Cys), member 5; Small-inducible cytokine A5; T cell-specific protein P228; T-cell specific protein p288; TCP228T-cell-specific protein RANTES;
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