Mannose Binding Lectin (MBL) also called mannose- or mannan-binding protein (MBP) is a member of the group of collectins. MBL is an oligomeric lectin that recognizes carbohydrates as mannose and N-acetylglucosamine on pathogens. MBL contains a cysteine rich, a collagen like and a carbohydrate recognition domain. It forms a complex with C1r/C1s like serine proteases designated MASPs that proteolytically cleave C4, C2 and C3. MBL is able to activate the complement pathway independent of the classical and alternative complement activation pathways. The MBL-MASP pathway (better known as the lectin pathway) is antibody and C1q-independent. MBL exhibits complement-dependent antibacterial activity and acts directly as an opsonic and therefore plays an important role in innate immunity. MBL is synthesized by hepatocytes and has been isolated from the liver or serum of various vertebrate species.
Flow cytometry, Frozen sections, Functional studies, Immuno assays
FC: cells were incubated with 1.0 μg/ml MBL in PBS containing 1%BSA (w/v), 0.01% NaN3 (w/v) and 10 mM CaCl2 for 30 min at 0°C. The cells were washed with the same buffer, and incubated with antibody clone 3E7, 6.5 μg/ml. After 30 min at 0°C, cells were washed and incubated with phycoerythrin (PE)-conjugated goat F(ab’)2 anti-mouse Ig (Ref.3).
IHC-F: Tissue sections embedded in ornithine carbamoyltransferase compound were frozen in acetone dry ice and immersed in Tris-buffered saline (TBS) three times for 5 minutes. Treatment in 0.5% hydrogen peroxide in methanol solution has been done to quench endogenous peroxidase activity (Ref.2).
FS: Pretreatment of sheep erythrocytes with antibody 3E7 suppressed hemolysis in a dose dependent manner (Ref.1).
Dilutions to be used depend on detection system applied. It is recommended that users test the reagent and determine their own optimal dilutions. The typical starting working dilution is 1:50. For functional studies, in vitro dilutions have to be optimized in user’s experimental setting.
Nonimmune normal serum or TBS