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Horse myeloperoxidase, MPO ELISA Kit
Overview
Size: 96T
Range: 1.25 ng/ml- 20 ng/ml
Sensitivity：0.1 ng/ml
Storage and Expiration: Store at 2-8℃ for 6 months.
Application: For quantitative detection of MPO in horse serum, plasma, tissue homogenate or cell culture supernatant.
Introduction
Myeloperoxidase (MPO) is a peroxidase enzyme that in humans is encoded by the MPO gene on chromosome 17. MPO is most abundantly expressed in neutrophil granulocytes (a subtype of white blood cells), and produces hypohalous acids to carry out their antimicrobial activity. It is a lysosomal protein stored in azurophilic granules of the neutrophil and released into the extracellular space during degranulation. MPO contains a calcium binding site with seven ligands, forming a pentagonal pyramid conformation. MPO is a member of the XPO subfamily of peroxidases and produces hypochlorous acid (HOCl) from hydrogen peroxide (H2O2) and chloride anion (Cl−) (or the equivalent from a non-chlorine halide) during the neutrophil's respiratory burst.
Principle of the Assay
This kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. The purified anti- MPO antibody was pre-coated onto 96-well plates. And the HRP conjugated anti- MPO antibody was used as detection antibodies. The standards, test samples and HRP conjugated detection antibody were added to the wells subsequently, mixed and incubated, then, unbound conjugates were washed away with wash buffer. TMB substrates (A & B) 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- horse MPO antibody
2. Standard: 0.3 ml × 6 tubes (0 ng/ml, 1.25 ng/ml, 2.5 ng/ml, 5 ng/ml, 10 ng/ml, 20 ng/ml)
3. Wash buffer (20×): 25 ml. Dilution: 1:20
4. Sample diluent buffer: 6 ml
5. HRP conjugated anti- horse MPO antibody (RTU): 10 ml
6. Stop solution: 6 ml
7. TMB substrate A: 6ml
8. TMB substrate B: 6ml
9. Plate sealer: 2
10. Hermetic bag: 1
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. Absorbent filter papers
8. Plastic or glass container with volume of above 1L
Other Information
Catalog No.: BYEK3449
Size: 96T
Range: 1.25 ng/ml- 20 ng/ml
Sensitivity：0.1 ng/ml
Storage and Expiration: Store at 2-8℃ for 6 months.
Application: For quantitative detection of MPO in horse serum, plasma, tissue homogenate or cell culture supernatant.
Introduction
Myeloperoxidase (MPO) is a peroxidase enzyme that in humans is encoded by the MPO gene on chromosome 17. MPO is most abundantly expressed in neutrophil granulocytes (a subtype of white blood cells), and produces hypohalous acids to carry out their antimicrobial activity. It is a lysosomal protein stored in azurophilic granules of the neutrophil and released into the extracellular space during degranulation. MPO contains a calcium binding site with seven ligands, forming a pentagonal pyramid conformation. MPO is a member of the XPO subfamily of peroxidases and produces hypochlorous acid (HOCl) from hydrogen peroxide (H2O2) and chloride anion (Cl−) (or the equivalent from a non-chlorine halide) during the neutrophil's respiratory burst.
Principle of the Assay
This kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. The purified anti- MPO antibody was pre-coated onto 96-well plates. And the HRP conjugated anti- MPO antibody was used as detection antibodies. The standards, test samples and HRP conjugated detection antibody were added to the wells subsequently, mixed and incubated, then, unbound conjugates were washed away with wash buffer. TMB substrates (A & B) 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- horse MPO antibody
2. Standard: 0.3 ml × 6 tubes (0 ng/ml, 1.25 ng/ml, 2.5 ng/ml, 5 ng/ml, 10 ng/ml, 20 ng/ml)
3. Wash buffer (20×): 25 ml. Dilution: 1:20
4. Sample diluent buffer: 6 ml
5. HRP conjugated anti- horse MPO antibody (RTU): 10 ml
6. Stop solution: 6 ml
7. TMB substrate A: 6ml
8. TMB substrate B: 6ml
9. Plate sealer: 2
10. Hermetic bag: 1
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. Absorbent filter papers
8. Plastic or glass container with volume of above 1L
Other Information
Please check datasheet for protocol.
