Anti-Mannan-binding lectin (human, MBL)

Mouse monoclonal antibody

Cat.No. HYB 131-01

Subclass IgG1/k
Presentation:

Preparation: Protein-A purified
Content: Available in 200 µL and 1 mL volumes, 1 mg/mL
Solvent: 0.01 M phosphate buffer, pH 7.4, containing 0.5 M NaCl and 15 mM sodium azide
Storage: In the dark at 4-8ºC

Antigen:

Mannan-binding lectin (MBL), also called mannose-binding lectin or protein, belongs to the C-type family of collectins, showing calcium-dependent binding to certain sugars (1). It consits of oligomers of triple-chain subunits and its binding and complement activating activities depend on its normal oligomerization. On binding to mannan-like microbial surface carbohydrates, MBL activates the complement system by means of its own lectin pathway, dependent on the MBL-associated serine proteases (MASPs). Because of the presence of different structural and promoter alleles in the population, 12% or more of the population have low concentrations (<50ng/mL) of normally oligomerized, functional MBL in plasma or serum (2)

Immunogen:

MBL purified from human donor plasma and adsorbed onto aluminum hydroxide.

Specificity:

HYB 131-01 is specific for MBL from human serum or plasma

Epitope specificity:

The epitope is on the head-neck region of the MBL protein chain. Prior binding of the antibody is thought to block binding to carbohydrate. The epitope specificity differs from that of HYB 131-10 and HYB 131-11.

Reactivity:

HYB 131-01 is selective for normally oligomerized MBL when used as detection antibody in sandwich ELISA with HYB 131-01 coat.

In Western blotting, HYB 131-01 reacts with human MBL in both its oligomerized state and as a single protein chain of 26 kDa.

HYB 131-01 is also well suited for immunohistochemistry on human tissue samples, frozen or paraffin embedded, from liver and brain.

Application:
Method Usability Dilution guideline References
ELISA Yes 1/20,000 3
Immunoblotting Yes 1/1000 4
Immunohistochemistry Yes 5
Application Remarks:

The dilution guideline for ELISA is based on use as detection antibody for antigen coated at 0.3 µg/mL together with human albumin 8.5 µg/mL. Suggested dilution when used as capture antibody in a sandwich ELISA is 5 µg/mL.

 
References:

1. Kawasaki N, Kawasaki T, Yamashina I (1983) Isolation and characterization of a mannan-binding protein from human serum. J Biochem (Tokyo) 94:937-947.

2. Steffensen R, Thiel S, Varming K, Jersild C, Jensenius JC (2000) Detection of structural gene mutations and promoter polymorphisms in the mannan-binding lectin (MBL) gene by polymerase chain reaction with sequence-specific primers. J Immunol Methods 241:33-42.

3. Garred P, Madsen MO, Kurtzhals JA, Lamm LU, Thiel S, Hey AS, Svejgaard A (1992) Diallellic polymorphism may explain variations of the blood concetration of mannan-binding protein in Eskimos, but not in black Africans. Eur J Immunogenet 19:403-412

4. Garred P, Larsen F, Madsen HO, Koch C (2003) Mannose-binding lectin deficiency - revisited. Mol Immunol 40:73-84

5. Holmskov UL (2000) Collectins and collectin receptors in innate immunity. APMIS Suppl 100:1-59.

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