Human Rantes ELISA Kit

Catalog No.: BEK1193

Size: 96T

Range: 15.6 pg/ml-1000 pg/ml

Sensitivity < 3 pg/ml

Storage and Expiration: Store at 2-8℃ for 6 months, or at -20℃ for 12 months. 

Application: For quantitative detection of Rantes in human serum, plasma, body fluids, tissue lysates or cell culture supernatants.

Introduction

Rantes, also known as Chemokine (C-C motif) ligand 5 (also CCL5), is a protein which in humans is encoded by the CCL5 gene, this chemokine has been localized to chromosome 17 in humans. Rantes is an 8kDa protein classified as a chemotactic cytokine or chemokine. It is chemotactic for T cells, eosinophils, and basophils. The Rantes plays an active role in recruiting leukocytes into inflammatory sites. With the help of particular cytokines (i.e., IL-2 and IFN-γ) that are released by T cells, CCL5 also induces the proliferation and activation of certain natural-killer (NK) cells to form CHAK (CC-Chemokine-activated killer) cells. It is also an HIV-suppressive factor released from CD8+ T cells. Recently, the RANTES protein has been engineered for in vivo production by Lactobacillus bacteria, and this solution is being developed into a possible HIV entry-inhibiting topical microbicide.

Principle of the Assay  

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti-Rantes polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti-Rantes polyclonal antibody was used as detection antibodies. The standards, test samples and biotin conjugated detection antibody were added to the wells subsequently, and wash with wash buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with wash buffer. TMB substrates were used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the Rantes amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of Rantes can be calculated.

Kit components

1. One 96-well plate pre-coated with anti-Human Rantes antibody
2. Lyophilized Human Rantes standards: 2 tubes (10 ng / tube)                     
3. Sample / Standard diluent buffer: 30ml
4. Biotin conjugated anti-Human Rantes antibody (Concentrated): 130μl. Dilution: 1:100
5. Antibody diluent buffer: 12ml
6. Avidin-Biotin-Peroxidase Complex (ABC) (Concentrated): 130μl. Dilution: 1:100
7. ABC diluent buffer: 12ml
8. TMB substrate: 10ml
9. Stop solution: 10ml
10. Wash buffer(25X): 30ml

Note: Reconstitute standards and test samples with Kit Component 3.

Material Required But Not Provided

1. 37℃ incubator
2. Microplate reader (wavelength: 450nm)
3. Precise pipette and disposable pipette tips
4. Automated plate washer
5. ELISA shaker
6. 1.5ml of Eppendorf tubes
7. Plate cover
8. Absorbent filter papers
9. Plastic or glass container with volume of above 1L