Name
MASP-2, Human, ELISA kit
Catalog nr
HK326
(lot number and expiry date are indicated on the label)
Short description
MASP-2 is a MBL associated serine protease. Mannan-binding lectin (MBL) and ficolins, in complex with MASPs, are capable of activating the complement system, thus mediating the destruction of infectious agents. After oligomerisation of MBL (or L-Ficolin) MASP-2 will be bound and activated. Activated MASP-2 cleaves C4 and C2 and is crucial for the activation of downstream complement components....
Size
2 x 96 det.
Application
Manual
Description
MASP-2 is a MBL associated serine protease. Mannan-binding lectin (MBL) and ficolins, in complex with MASPs, are capable of activating the complement system, thus mediating the destruction of infectious agents. After oligomerisation of MBL (or L-Ficolin) MASP-2 will be bound and activated. Activated MASP-2 cleaves C4 and C2 and is crucial for the activation of downstream complement components. MASP-2 deficiency will result in an increased susceptibility for infections (e.g. leukemia patients in chemotherapy are at high risk of serious infections). Even more so then MBL deficiency, because MASP-2 is also required for complement activation by H- and L-ficolin. The ELISA detects MASP-2 in serum and plasma. The MASP-2 levels in plasma from healthy individuals range from 170 to 1196 ng/ml.
Application
The human MASP-2 ELISA has been developed for the quantitative measurement of natural human MASP-2 in cell culture medium, plasma and serum. In serum or plasma samples human MASP-2 can be measured accurately if serum or plasma samples are diluted at least 4 times. Most reliable results are obtained if EDTA plasma is used.
Features
- Minimum concentration which can be measured is 1.6 ng/ml human MASP-2.
- Measurable concentration range of 1.6-100 ng/ml.
- Working volume of 100 µl/well.
Typical standard curve
Principle
- The human MASP-2 ELISA is a ready-to-use solid-phase enzyme-linked immunosorbent assay based on the sandwich principle which a working time of 3½ hours.
- The efficient format of 2 plates with twelve disposable 8-well strips allow 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 human MASP-2.
- 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 human MASP-2 standards (log).
- The human MASP-2 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 standard is stable for 1 month at 4°C.
Recovery
Normal human blood samples (plasma), containing baseline levels of human MASP-2, were spiked with recombinant MASP-2 in concentrations of 25 and 2.5 ng/ml. Samples with and without human MASP-2 were incubated for 1 hour at room temperature. Samples were measured using the ELISA. Values for human MASP-2 ranged between 123% and 87% (mean 99.8%).
Linearity
The linearity of the assay was determined by serially diluting a sample containing 328 ng/ml human MASP-2. The diluted samples were measured in the assay. The line obtained a slope of 1.01 and a correlation coefficient of 0.999. 
References
- Moeller-Kritensen, M et al; Levels of mannan-binding lectin-associated serine protease-2 in healthy individuals. J Immunol Meth 2003, 282: 159
- Schlapbach, L et al; Deficiency of mannose-binding lectin-associated serine protease-2 associated with increase risk of fever and neutropenia in pediatric cancer patients. Ped Infect Dis 2007, 26: 989
Precautions
For research use only. The kit is to be used for research purposes only, not for clinical or diagnostic use. The MASP-2 ELISA is Patent protected by patent 09/874,238 and PCT/DK2004/000338. License rights on the Patents are acquired from Natimmune S/A, Denmark. 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
- Moeller-Kritensen, M et al; Levels of mannan-binding lectin-associated serine protease-2 in healthy individuals. J Immunol Meth 2003, 282: 159
- Schlapbach, L et al; Deficiency of mannose-binding lectin-associated serine protease-2 associated with increase risk of fever and neutropenia in pediatric cancer patients. Ped Infect Dis 2007, 26: 989
Scientific info
Higher cord blood MASP-2 levels: a risk factor for NEC
Higher MASP-2 levels may favor complement-mediated inflammation and could thereby predispose to necrotizing enterocolitis (NEC). Determination of MASP-2 with the MASP-2 ELISA (Cat. # HK326) may help to identify infants at increased risk of NEC.
Schlapbach, L et al; Higher cord blood levels of mannose-binding lectin-associated serine protease-2 in infants with necrotising enterocolitis. Pediatr Res 2008
Higher MASP-2 levels may favor complement-mediated inflammation and could thereby predispose to necrotizing enterocolitis (NEC). Determination of MASP-2 with the MASP-2 ELISA (Cat. # HK326) may help to identify infants at increased risk of NEC.
NEC is a major cause of morbidity and mortality in premature infants. Unfortunately, the exact pathophysiological mechanisms underlying NEC remain unidentified and identification of infants at risk for NEC is a challenge.
The lectin pathway of the complement system may be involved in NEC pathogenesis by generation of potentially harmful inflammatory mediators. Mannose-binding lectin-associated serine protease-2 (MASP-2) represents the common pathway for both mannose binding lectin (MBL) and ficolins.
Schlapbach and co-workers investigated the correlation between cord blood concentrations of MBL and MASP-2 and later development of NEC. Higher cord blood MASP-2 (but not MBL) levels were significantly associated with an increased risk of NEC in multivariate analysis when compared to controls.
MASP-2 cord blood concentrations were extremely low in most neonates compared to values reported from adults and children. MASP-2 deficiency, resulting in extremely low MASP-2 concentrations, suggests that the lectin pathway of complement activation is often not fully functional at birth. This may represent a protective mechanism against excessive proinflammatory stimuli during the neonatal period.
Schlapbach, L et al; Higher cord blood levels of mannose-binding lectin-associated serine protease-2 in infants with necrotising enterocolitis. Pediatr Res 2008
MSDS
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