Human MDC ELISA Kit

Catalog No.: BEK1148

Size: 96T

Range: 31.2 pg/ml-2000 pg/ml

Sensitivity < 1 pg/ml

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

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

Introduction

Macrophage-derived chemokine (MDC), also called Chemokine, cc motif, ligand 22 (CCL22) or Small inducible cytokine subfamily A, member 22 (SCY22), is a protein that in humans is encoded by the CCL22 gene. The gene is located in human chromosome 16 in a cluster with other chemokines called CX3CL1 and CCL17. It is secreted by dendritic cells and macrophages, and elicits its effects on its target cells by interacting with cell surface chemokine receptors such as CCR4. HTLV-1-induced CCL22 causes the high frequency of FOXP3-positive cells observed in HTLV-1 infection and that FOXP3-positive cells may both retard the progression of ATLL and HTLV-1-associated inflammatory diseases and contribute to the immune suppression seen in HTLV-1 infection.

Principle of the Assay  

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti- MDC polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti- MDC 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 MDC amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of MDC can be calculated.

Kit components

1. One 96-well plate pre-coated with anti-Human MDC antibody
2. Lyophilized Human MDC standards: 2 tubes (10 ng / tube)                     
3. Sample / Standard diluent buffer: 30ml
4. Biotin conjugated anti-Human MDC 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