Name
MPO, Rat, ELISA kit
Catalog nr
HK105-01
(lot number and expiry date are indicated on the label)
Short description
Myeloperoxidase (MPO) is a glycoprotein with an 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...
Size
1 x 96 det.
Application
Manual
Description
Myeloperoxidase (MPO) is a glycoprotein with an 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. MPO is released in the extra cellular medium where hypochlorous acid leads to chlorination of proteins leading to products as 3-chlorotyrosine. Furthermore it leads to oxidation of low density lipoprotein (LDL) and apolipoprotein A-I, the primary protein of HDL resulting in the disruption of HDL functions as cholesterol efflux. 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 rat MPO elisa is not influenced by inhibitors of the enzyme activity.
Application
The rat MPO ELISA has been developed for the quantitative measurement of natural and recombinant rat MPO in cell culture medium, plasma and tissue homogenates. In plasma samples MPO can be measured accurately if plasma samples are diluted at least 4 times. Most reliable results are obtained if EDTA plasma is used. Please be aware that rat MPO is released from neutrophils into serum in the process of blood coagulation. This will lead to false positive and difficult to interpret results of serum samples. Therefore it is advised to use 'careful plasma'.
Features
- Minimum concentration which can be measured is 3.9 ng/ml rat MPO.
- Measurable concentration range of 3.9-250 ng/ml.
- Working volume of 100 µl/well.
Typical standard curve
Principle
- The rat 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 captured by a solid bound specific antibody.
- Biotinylated tracer antibody will bind to captured rat 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 citric 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 rat MPO standards (log).
- The rat MPO concentration of samples, which are run concurrently with the standards, can be determined from the standard curve.
Storage and stability
Product should be stored at 4°C. Under recommended storage conditions, product is stable for at least six months. After reconstitution the reagents are stable for 1 month if stored at 2-8°C.
Recovery
Normal rat blood samples (plasma), were spiked with rat MPO in concentrations of 100 ng/ml. Samples with and without rat MPO were incubated for 1 hour at room temperature. Samples were measured using the ELISA. Recovery values for rat MPO ranged between 108% and 120% (mean 114 %).
Linearity
The linearity of the assay was determined by serially diluting a serum sample containing 50 ng/ml rat MPO. The diluted samples were measured in the assay. The line obtained a slope of 0.90 and a correlation coefficient of 0.999. 
References
1. Roelofs, L et al; Tissue-type plasminogen activator modulates inflammatory responses and renal function in ischemia reperfusion injury. J Am Soc Nephrol 2006, 17: 131
Precautions
For research use only. Not for use in or on humans or animals or for diagnostics. It is the responsibility of the user to comply with all local/state and Federal rules in the use of this product. Hycult Biotech is not responsible for any patent infringements that might result with the use of or derivation of this product.
Also available
References
- Roelofs, L et al; Tissue-type plasminogen activator modulates inflammatory responses and renal function in ischemia reperfusion injury. J Am Soc Nephrol 2006, 17: 131
Scientific info
Rat Myeloperoxidase (MPO) assay for tissue neutrophil infiltration
Special features:
• Reliable and fast technique based on highly specific antibodies for MPO
• Not influenced by tissue enzyme inhibitors
• No time consuming histology and microscopic cell counting
• Protocol for tissue homogenization included
MPO and tissue enzyme inhibitors
The classical enzymatic assay for MPO quantification in tissue requires the blockade of inhibitory factors. Especially organs like kidney and liver contain large amounts of inhibitors that disturb the enzymatic MPO assay in such tissues. The Hbt assay is an ELISA based on highly specific MPO antibodies. This method is not influenced by tissue enzyme inhibitors and allows accurate and easy detection of MPO present in infiltrating neutrophils. Furthermore, the assay can be used for quantification of MPO in plasma, e.g. after LPS administration, during inflammation and for quantification of MPO during in vitro release from neutrophils.
MPO
Myeloperoxidase (MPO) is an alpha2beta2 heteromultimer glycoprotein expressed in all cells of the myeloid linage. MPO is abundantly present in the azurophilic granules of polymorphonuclear neutrophils from where it is released upon neutrophil activation. MPO is an important enzyme used during phagocytic lysis of engulfed foreign particles. This organism’s defense mechanism takes part through production of hypochlorous acid (HOCl), a potent oxidant.
Schematic representation of MPO release from azurophilic granules of a polymorphonuclear neutrophil
MPO and disease
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.
Selected reading
1. Gultekin, F et al; Leptin treatment ameliorates acute lung injury in rats with cerulein-induced acute pancreatitis. World J Gastroenterol, 2007, 13: 2932
2. Roelofs, J et al; Tissue-type plasminogen activator modulates inflammatory responses and renal function in ischemia reperfusion injury. J Am Soc Nephrol, 2006, 17: 131
Special features:
• Reliable and fast technique based on highly specific antibodies for MPO
• Not influenced by tissue enzyme inhibitors
• No time consuming histology and microscopic cell counting
• Protocol for tissue homogenization included
MPO and tissue enzyme inhibitors
The classical enzymatic assay for MPO quantification in tissue requires the blockade of inhibitory factors. Especially organs like kidney and liver contain large amounts of inhibitors that disturb the enzymatic MPO assay in such tissues. The Hbt assay is an ELISA based on highly specific MPO antibodies. This method is not influenced by tissue enzyme inhibitors and allows accurate and easy detection of MPO present in infiltrating neutrophils. Furthermore, the assay can be used for quantification of MPO in plasma, e.g. after LPS administration, during inflammation and for quantification of MPO during in vitro release from neutrophils.
MPO
Myeloperoxidase (MPO) is an alpha2beta2 heteromultimer glycoprotein expressed in all cells of the myeloid linage. MPO is abundantly present in the azurophilic granules of polymorphonuclear neutrophils from where it is released upon neutrophil activation. MPO is an important enzyme used during phagocytic lysis of engulfed foreign particles. This organism’s defense mechanism takes part through production of hypochlorous acid (HOCl), a potent oxidant.
Schematic representation of MPO release from azurophilic granules of a polymorphonuclear neutrophil
MPO and disease
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.
Selected reading
1. Gultekin, F et al; Leptin treatment ameliorates acute lung injury in rats with cerulein-induced acute pancreatitis. World J Gastroenterol, 2007, 13: 2932
2. Roelofs, J et al; Tissue-type plasminogen activator modulates inflammatory responses and renal function in ischemia reperfusion injury. J Am Soc Nephrol, 2006, 17: 131
MSDS
| Related products | Cat # | |
|---|---|---|
| MPO, Human, ELISA kit | HK324-02 | |
| MPO, Mouse, ELISA kit | HK210-02 | |