Human Myeloperoxidase / MPO ELISA Kit

Catalog No.: BEK1165

Size: 96T

Range: 312 pg/ml-20,000 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 MPO in human serum, body fluids, tissue lysates or cell culture supernatants.

Introduction

Myeloperoxidase (MPO) is a peroxidase enzyme that in humans is encoded by the MPO gene which contains 12 exons and spans about 13 kb. Myeloperoxidase is most abundantly expressed in neutrophil granulocytes. It is a lysosomal protein stored in azurophilic granules of the neutrophil. The 150-kDa MPO protein is a dimer consisting of two 15-kDa light chains and two variable-weight glycosylated heavy chains bound to a prosthetic heme group. MPO produces hypochlorous acid (HOCl) from hydrogen peroxide (H2O2) and chloride anion (Cl-) during the neutrophil's respiratory burst. It requires heme as a cofactor. Furthermore, it oxidizes tyrosine to tyrosyl radical using hydrogen peroxide as an oxidizing agent.

Principle of the Assay  

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

Kit components

1. One 96-well plate pre-coated with anti-Human MPO antibody
2. Lyophilized Human MPO standards: 2 tubes (20 ng / tube)                     
3. Sample / Standard diluent buffer: 30ml
4. Biotin conjugated anti-Human MPO 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