Name
MPO, Human, ELISA kit
Catalog nr
HK324-01
(lot number and expiry date are indicated on the label)
Short description
Myeloperoxidase (MPO) is a glycoprotein with a alpha2beta2 heteromultimer expressed in all cells of the myeloid linage. MPO is abundantly present in azurophilic granules of polymorphonuclear neutrophils. It is an important enzyme used during phagocytic lysis of engulfed foreign particles which takes part in the defense of the organism through production of hypochlorous acid (HOCl), a potent o...
Size
1 x 96 det.
Application
Manual
Description
Myeloperoxidase (MPO) is a glycoprotein with a alpha2beta2 heteromultimer expressed in all cells of the myeloid linage. MPO is abundantly present in azurophilic granules of polymorphonuclear neutrophils. It is an important enzyme used during phagocytic lysis of engulfed foreign particles which takes part in the defense of the organism through production of hypochlorous acid (HOCl), a potent oxidant. MPO is rapidly released by activated polymorphonuclear neutrophils. Involvement of MPO has been described in numerous diseases such as atherosclerosis, lung cancer, Alzheimer's disease and multiple sclerosis. Autoimmune antibodies to MPO are involved in Wegener’s disease. Since the discovery of MPO deficiency, initially regarded as rare and restricted to patients suffering from severe infections, MPO has attracted more clinical attention.
The classical MPO assay is an enzymatic assay for activity of MPO. This classical MPO assay is hampered by the presence of inhibitory compounds in tissue homogenates and plasma. In this type of assays spiking often gives unreliable results. The human MPO ELISA is not influenced by inhibitors of the enzyme activity.
Cross Reactivity
Potential cross-reacting proteins detected in the human MPO ELISA:
|
Cross reactant |
Reactivity |
|
Mouse MPO |
negative |
|
Rat MPO |
negative |
|
Rabbit MPO |
negative |
|
Bovine MPO |
negative |
|
Sheep MPO |
negative |
|
Swine MPO |
negative |
|
Horse MPO |
negative |
Species
Human
Features
- Working time of 3½ hours.
- Minimum concentration which can be measured is 0.4 ng/ml.
- Measurable concentration range of 0.4 to 100 ng/ml.
- Working volume of 100 µl/well.
Typical standard curve
Principle
- The human MPO ELISA is a ready-to-use solid-phase enzyme-linked immunosorbent assay based on the sandwich principle with a working time of 3½ hours.
- The efficient format of a plate with twelve disposable 8-well strips allows free choice of batch size for the assay.
- Samples and standards are incubated in microtiter wells coated with antibodies recognizing human MPO.
- Biotinylated tracer antibody will bind to captured human MPO.
- Streptavidin-peroxidase conjugate will bind to the biotinylated tracer antibody.
- Streptavidin-peroxidase conjugate will react with the substrate, tetramethylbenzidine (TMB).
- The enzyme reaction is stopped by the addition of oxalic acid.
- The absorbance at 450 nm is measured with a spectrophotometer. A standard curve is obtained by plotting the absorbance (linear) versus the corresponding concentrations of the human MPO standards (log).
- The human MPO concentration of samples, which are run concurrently with the standards, can be determined from the standard curve.
Use
The human MPO ELISA kit is to be used for the in vitro quantitative determination of human MPO in plasma, sputum and cell culture supernatant samples. This kit is intended for laboratory research use only and is not for use in diagnostic or therapeutic procedures.
Storage and stability
- Upon receipt, store individual components at 2 - 8°C. Do not freeze.
- Do not use components beyond the expiration date printed on the kit label.
- The standard, tracer and streptavidin-peroxidase are stable in lyophilized form until the expiration date indicated on the kit label, if stored at 2 - 8°C.
- The exact concentration of the standard is indicated on the label of the vial and the certificate of quality control.
- Once reconstituted the standard is stable for 24 hours, if stored at 2 - 8°C. For longer stability we recommend to store aliquots at –20°C. Stored at –20°C the standard will be stable for 1 month.
- Once reconstituted, tracer and streptavidin-peroxidase are stable for 1 month if stored at 2 - 8°C.
- Upon receipt, foil pouch around the plate should be vacuum-sealed and unpunctured. Any irregularities to aforementioned conditions may influence plate performance in the assay.
- Return unused strips immediately to the foil pouch containing the desiccant pack and reseal along the entire edge of the zip-seal. Quality guaranteed until expiration date if stored at 2 - 8°C.
References
- Leclercq A et al; Involvement of intraplaque hemorrhage in atherothombosis evolution via neutrophil protease enrichment. J Leukoc Biol 2007, 82: 1420
- Martin-Ventura J et al ; Low plasma levels of HSP70 in patients with carotid atherosclerosis are associated with increased levels of proteolytic markers of neutrophil activation. Atherosclerosis 2007, 194: 334
- Puklo, M et al; Analysis of neutrophil-derived antimicrobial peptides in gingival crevicular fluid suggests importance of cathelicidin LL-37 in the innate immune response against periodontogenic bacteria. Oral Microbiol Immunol 2008, 23: 328
- Haan de, J et al; Postshock intervention with high-lipid enteral nutrition reduces inflammation and tissue damage. Ann Surg 2008, 248: 842
References
- Leclercq A et al; Involvement of intraplaque hemorrhage in atherothombosis evolution via neutrophil protease enrichment. J Leukoc Biol 2007, 82: 1420
- Martin-Ventura J et al ; Low plasma levels of HSP70 in patients with carotid atherosclerosis are associated with increased levels of proteolytic markers of neutrophil activation. Atherosclerosis 2007, 194: 334
- Puklo, M et al; Analysis of neutrophil-derived antimicrobial peptides in gingival crevicular fluid suggests importance of cathelicidin LL-37 in the innate immune response against periodontogenic bacteria. Oral Microbiol Immunol 2008, 23: 328
- Haan de, J et al; Postshock intervention with high-lipid enteral nutrition reduces inflammation and tissue damage. Ann Surg 2008, 248: 842
MSDS
| Related products | Cat # | |
|---|---|---|
| LL-37, Human, ELISA kit | HK321-02 | |
| Elastase, Human, ELISA kit | HK319-02 | |
| Calprotectin, Human, ELISA kit | HK325-02 | |
| HNP1-3, Human, ELISA kit | HK317-01 | |
| Lactoferrin, Human, ELISA kit | HK329 | |
| MPO, Mouse, ELISA kit | HK210-02 | |
| MPO, Rat, ELISA kit | HK105-02 | |