Human MIP-1α ELISA Kit

Catalog No.: BEK1152

Size: 96T

Range: 15.6 pg/ml - 1000 pg/ml

Sensitivity < 10 pg/ml

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

Application: For quantitative detection of MIP-1α in human serum, plasma, body fluids, tissue lysates or cell culture supernatants.

Introduction

Macrophage inflammatory protein-1(MIP-1a), also known as Chemokine (C-C motif) ligand 3(CCL3) or LD78, is a cytokine belonging to the CC chemokine family. The LD78 gene chromosome 17q21.1-q21.3. It is involved in the acute inflammatory state in the recruitment and activation of polymorphonuclear leukocytes. Sherry et al. (1988) demonstrated 2 protein components of MIP1, called by them alpha and beta. MIP-1a is an important mediator of virus-induced inflammation in vivo, and also an important second signal for mast cell degranulation in the conjunctiva and for acute-phase disease, possibly through interaction with CCR1, its chemokine receptor.

Principle of the Assay  

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

Kit components

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