Name
MBL, Human, ELISA kit
Catalog nr
HK323-01
(lot number and expiry date are indicated on the label)
Short description
Mannose Binding Lectin (MBL) is a C-type lectin that is an important element in innate immunity. MBL belongs to the collectin family of proteins that consists of a collagen-like domain and a carbohydrate recognition domain (CRD). Through this, MBL recognizes carbohydrates (e.g. mannose and N-acetylglucosamine) on pathogens. MBL forms several sizes of oligomers that are composed of subunits of...
Size
1 x 96 det.
Application
Manual
Description
Mannose Binding Lectin (MBL) is a C-type lectin that is an important element in innate immunity. MBL belongs to the collectin family of proteins that consists of a collagen-like domain and a carbohydrate recognition domain (CRD). Through this, MBL recognizes carbohydrates (e.g. mannose and N-acetylglucosamine) on pathogens. MBL forms several sizes of oligomers that are composed of subunits of three identical 32 kDa polypeptides. MBL is a pattern recognition receptor, and upon recognition of the infectious agent, MBL triggers the activation of the lectin-complement pathway through attached C1r/C1s-like serine proteases, designated MASP. The MBL-MASP complex proteolytically cleaves C4, C2 and C3. The lectin pathway is antibody and C1q-independent and is, therefore, independent of the classical and alternative complement activation pathway. MBL is synthesized by hepatocytes and has been isolated from the liver or serum of several vertebrate species. Normal human plasma contains MBL concentrations ranging from 10 to 5,000 ng/ml. Up to 12% of healthy Caucasian blood donors have MBL concentrations below 100 ng/ml. Low plasma concentrations have been associated with an inherited defect in opsonization. The MBL concentration is enhanced in infectious diseases. Measuring of MBL is indicated in recurrent infections (especially in children), primary/secondary immunodeficiencies, artherosclerosis/coronary heart disease, cystic fibrosis, transplantation, autoimmune diseases (SLE/Rheumatoid arthritis) and habitual abortion. MBL measurement is not affected by the presence of antibodies against mannan. In the human MBL assay, any influence of the classical pathway has been eliminated by using a special MBL-binding buffer, which inhibits the binding of C1q to immunocomplexes and disrupts the C1 complex, while leaving the function of the MBL complex intact.
Application
The human MBL (Lectin assay) ELISA has been developed for the quantitative measurement of natural functional, mannan binding, human MBL in serum, plasma and culture medium. In serum or plasma samples, MBL 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 0.41 ng/ml MBL.
- Measurable concentration range of 0.41-100 ng/ml.
- Working volume of 100 µl/well.
Typical standard curve
Principle
- The human MBL ELISA is a ready-to-use solid-phase enzyme-linked immunosorbent assay based on the sandwich principle with a working time of 4 hours.
- The efficient format of a plate with twelve disposable 8-well strips allows free choice of batch size for the assay.
- After activation, samples and standards are captured by solid bound mannan.
- Biotinylated tracer antibody will bind to captured MBL.
- 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 MBL standards (log).
- The MBL concentration of samples, which are run concurrently with the standards, can be determined from the standard curve.
Aliases
Mannose- or mannan-binding protein (MBP)
Storage and stability
Product should be stored at 4°C. Under recommended storage conditions, product is stable for at least six months. Aliquots of the standard should be stored at -20°C. After reconstitution, all reagents but the standard are stable for 1 month if stored at 2-8°C.
References
- Kirkpatrick, B et al; Serum mannose-binding lectin deficiency is associated with cryptosporidiosis in young Haitian children. Clin Infect Dis 2006, 43: 289
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
- Kirkpatrick, B et al; Serum mannose-binding lectin deficiency is associated with cryptosporidiosis in young Haitian children. Clin Infect Dis 2006, 43: 289
Scientific info
Cardiac surgery patients deficient in mannose binding lectin are protected against multiple organ dysfunction syndrome.
Systemic inflammatory responses after cardiac surgery can evolve to multiple organ dysfunction syndrome (MODS). The outcome of cardiac surgery is therefore closely related with the severity of MODS and the development of severe infections.
One of the components of the inflammatory response that is activated during cardiac surgery is the lectin pathway of complement. This pathway can be triggered by binding of certain surface carbohydrates on microorganisms to mannose-binding lectin (MBL). In the April number of Transfusion researchers evaluate a relation between cardiac surgery, blood transfusions and pre- and post-operative MBL levels. Preoperative serum MBL levels of patients were categorized as high (> 400 ng/ml), low (≤ 400 ng/ml) or MBL deficient (≤ 80 ng/ml), as measured by a sandwich ELISA based on monoclonal anti-MBL antibody 3E7.
No association was observed between high or low pre- or postsurgery MBL levels and postoperative infections after cardiac surgery. However, patients with an obvious MBL deficiency before surgery, were protected against the development of MODS. Deficiency of MBL may preclude activation of the complement system and induction of the systemic inflammatory response upon ischemia and reperfusion injury. In conclusion, sustained MBL deficiency may be a favorable status for patients undergoing cardiac surgery.
References
1. Bilgin, Y et al; Mannose-binding lectin is involved in multiple organ dysfunction syndrome after cardiac surgery: effects of blood transfusions. Transfusion, 2008, 48: 601
2. Matsushita, M et al; Human mannose-binding protein is identical to a component of Ra-reactive factor. Biochem Biophys Res Commun 1992, 183: 645
Systemic inflammatory responses after cardiac surgery can evolve to multiple organ dysfunction syndrome (MODS). The outcome of cardiac surgery is therefore closely related with the severity of MODS and the development of severe infections.
One of the components of the inflammatory response that is activated during cardiac surgery is the lectin pathway of complement. This pathway can be triggered by binding of certain surface carbohydrates on microorganisms to mannose-binding lectin (MBL). In the April number of Transfusion researchers evaluate a relation between cardiac surgery, blood transfusions and pre- and post-operative MBL levels. Preoperative serum MBL levels of patients were categorized as high (> 400 ng/ml), low (≤ 400 ng/ml) or MBL deficient (≤ 80 ng/ml), as measured by a sandwich ELISA based on monoclonal anti-MBL antibody 3E7.
No association was observed between high or low pre- or postsurgery MBL levels and postoperative infections after cardiac surgery. However, patients with an obvious MBL deficiency before surgery, were protected against the development of MODS. Deficiency of MBL may preclude activation of the complement system and induction of the systemic inflammatory response upon ischemia and reperfusion injury. In conclusion, sustained MBL deficiency may be a favorable status for patients undergoing cardiac surgery.
References
1. Bilgin, Y et al; Mannose-binding lectin is involved in multiple organ dysfunction syndrome after cardiac surgery: effects of blood transfusions. Transfusion, 2008, 48: 601
2. Matsushita, M et al; Human mannose-binding protein is identical to a component of Ra-reactive factor. Biochem Biophys Res Commun 1992, 183: 645
MBL deficiency in preterm and very light neonates
Low MBL levels at birth are associated with an increased risk of early-onset sepsis, culture-proven sepsis and pneumonia during the first month of life. Even neonates with a wild-type MBL-2 genotype may be deficient in MBL at birth. Therefore, detection of MBL-deficiency at birth should be based on actual MBL plasma levels rather than on MBL-2 genotype.
Mannose-binding lectin (MBL) is a circulating pattern recognition molecule playing an important role in the innate immune defense. It activates the lectin pathway of the complement system. Circulating MBL plasma levels vary between 0 and 10 µg/ml. These variations are correlated with mutations in the structural and promoter regions of the MBL-2 gene that lead to reduced MBL concentrations.
Babies with MBL levels below 400 ng/ml are more likely to suffer from sepsis, regardless of their genotype. In neonates, regardless of their MBL-2 gene genotype, a very low birth weight (less than 1000g) or gestation of less than 28 weeks increased the risk of sepsis to 70%. Whereas, in neonates with MBL levels above 400 ng/ml, the risk of sepsis was less than 50%.
References:
1. Dzwonek, A et al; The role of mannose-binding lectin in susceptibility to infection in preterm neonates. Pediatr Res 2008, 63: 680
2. Frakking, F et al; Low mannose-binding lectin (MBL) levels in neonates with pneumonia and sepsis. Clin Exp Immunol 2007, 150: 255
3. Frakking, F et al; High prevalence of mannose-binding lectin (MBL) deficiency in premature neonates. Clin Exp Immunol 2006, 145: 5
MSDS
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