Rantes ELISA Kit (CCL5)

INTENDED USE

Human Rantes ELISA kit is intended for measuring in vitro quantitative levels of regulated on activation normal T cell expressed and secreted (rantes)(CCL5) in human biological fluids, serum, tissue homogenates and plasma (citrate, EDTA, heparin). This assay has a minimum detection limit of 18.75 pg/ml.

BACKGROUND

Rantes is also known as Chemokine (C-C motif) ligand 5 (CCL5), is essentially a natural ligand for the chemokine receptor CCR5 and has the ability to potently suppress the in vitro replication of R5 strains of HIV-1, that is using CCR5 as a co-receptor. It is chemotactic for basophils, eosinophils and T cells, it also plays a vital role in the recruitment of into inflammatory sites. The gene product is predicted to be 10 kDa and after cleavage of the signal peptide, approx. 8 kDa. From the 68 residues remaining, where 4 are cysteines and none are N-linked glycosylated.

Rantes is usually expressed by cultured T cell lines that are growth factor dependent and antigen specific. There is evidence that it can induce the activation and proliferation of a number of natural-killer cells to form CHAK Chemokine-activated killer) cells with the aid of particular cytokines (in particular IL-2 and IFN-γ). CCL5 is also known as a HIV-suppressive factor that can be released from CD8+ T cells. Peripheral blood mononuclear cells or CD4(+) lymphocytes that are obtained from different individuals have been found to display a wide variations in their ability to secrete rantes.

CONTENT

All reagents supplied need to be stored at 2 °C – 8 °C, unopened reagents will retain reactivity until expiration date. Do not use reagents beyond this date.

• One 96-Well Plate: Pre-coated with anti-human Rantes antibody.
• Standards: Concentrations 0, 31.25, 62.5, 125, 250, 500, 1000, 2000 pg/ml, made up from lyophilized recombinant.
• Sample/Standard Dilution Buffer.
• Biotinylated-Labelled Antibody.
• Antibody Dilution Buffer.
• HRP-Streptavidin Conjugate (SABC).
• SABC Dilution Buffer.
• TMB Substrate.
• Wash Buffer (25x).
• Plate Sealer.
• Product Instructions.

SENSITIVITY

The minimum detection sensitivity level of rantes (CCL5) using this rantes ELISA kit was 18.75 pg/ml. The dynamic assay range for this kit is 31.25 – 2,000 pg/ml.

ASSAY CHARACTERISTICS

– Specificity: Highly specific for CCL5, no cross reactivity or interference between CCL5 and analogues was detected.
– Recovery: Serum (86 – 101%), EDTA Plasma (88 – 104%), Heparin Plasma (88 – 100%).
– Linearity: Serum (88 – 104%), EDTA Plasma (93 – 101%), Heparin Plasma (82 – 89%).
– Precison Intra-Assay: CV < 8%.
– Precison Inter-Assay: CV < 10%.
– Stability: Less than 10%.

REFERENCES

1. RANTES: a versatile and controversial chemokine. Trends Immunol. (2001) 22 (2): 83-7. Review. Appay V. and Rowland-Jones S.L.
2. Angiogenic properties of the chemokine RANTES/CCL5. Biochem Soc Trans. (2011) 39 (6): 1649-53. Review. Suffee N., et al.
3. The inflammatory chemokine CCL5 and cancer progression. Mediators Inflamm. (2014) 2014: 292376. Review. Aldinucci D. and Colombatti A.
4. MCP-1 and RANTES are mediators of acute and chronic inflammation. Allergy Asthma Proc. (2001) 22 (3): 133-7. Review. Conti P. and DiGioacchino M.

ADDITIONAL INFORMATION

• Full Name: Rantes ELISA Kit (CCL5)
• Reactivity: Human
• Sample Type: Tissue Homogenates, Serum, Plasma, Other Biological Fluids
• Sensitivity: 18.75 pg/ml

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