Human MCP-1 ELISA Kit

Catalog No.: BEK1142

Size: 96T

Range: 15.6 pg/ml-1000 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 MCP-1 in Human serum, plasma, body fluids, tissue lysates or cell culture supernatants.

Introduction

Monocyte chemotactic protein-1(MCP-1), known as chemmkine(C-C motif) ligand 2(CCL2) or small inducible cytokine A2, is a small cytokine belonging to the CC chemokine family. The CCL2 gene is located on chromosome 17 (17q11.2-q21.1) in humans, and spans 1,927 bases. It has three exons and two introns, and produced as a protein precursor containing signal peptide of 23 amino acids and a mature peptide of 76 amino acids. The predicted weight is 11.025 kiloDaltons (kDa). MCP-1 has been implicated in the pathogenesis of diseases characterized by monocytic infiltrates, like psoriasis, rheumatoid arthritis and atherosclerosis. It induces amylin expression through ERK1/ ERK2/ JNK-AP1 and NF-κB related signaling pathways independent of CCR2. Amylin upregulation by CCL2 may contribute to elevation of plasma amylin in obesity and insulin resistance.

Principle of the Assay 

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti-MCP-1 polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti-MCP-1 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 MCP-1 amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of MCP-1 can be calculated.

Kit components

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