HBT ELISA TEST KIT FOR MOUSE MBL-C The Hbt mouse MBL-C test kit has been developed for the quantitative measurement of natural mouse MBL-C in serum, plasma and culture medium. Mannose Binding Lectin (MBL) also called mannose- or mannan-binding protein (MBP) is a member of the collectins and an important element in innate immunity. 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 MASP 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 anti-bacterial 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 and serum of several vertebrate species. Only one form of human MBL has been characterized, while two forms are found in rhesus monkeys, rabbits, rats and mice. The murine forms are known as MBL-A and MBL-C. The MBL-C concentrations in serum are about 6-fold higher compared to that of MBL-A. MBL-A, but not MBL-C was found to be an acute phase protein in casein and LPS-injection models. MBL-C exists in higher oligomeric forms than MBL-A. In plasma of healthy mice MBL-C concentrations range from 16 to 118 Âug/ml. In infectious diseases the MBL-C concentration was not found to increase significantly. PRINCIPLE OF THE TEST The Hbt mouse MBL-C Elisa test kit is a solid-phase enzyme-linked immunosorbent assay based on the sandwich principle. Samples and standards are incubated in microtiter wells coated with antibodies recognizing mouse MBL-C. During this incubation MBL-C is captured by the solid bound antibody. Unbound material present in the sample is removed by washing. Biotinylated antibody (tracer) to mouse MBL-C is added to the wells. If MBL-C was present in the sample, the tracer antibodies will bind to the captured MBL-C. Excess tracer is removed by washing. Streptavidin-peroxidase conjugate is applied to the wells, this conjugate reacts specifically with the biotinylated tracer antibody. Excess streptavidin-peroxidase conjugate is removed by washing and substrate, tetramethylbenzidine (TMB) is added to the wells. Color develops proportionally to the amount of MBL-C present in the sample. The enzyme reaction is stopped by the addition of citric acid and the absorbance at 450 nm is measured with a spectrophotometer. A standard curve is obtained by plotting the absorbance versus the corresponding concentrations of the standards. The MBL-C concentration of samples with unknown concentrations, which are run concurrently with the standards, can be determined from the standard curve. SPECIAL FEATURES OF THE KIT - Ready-to-use (ie. pre-coated microwells) - High reproducibility. - High specificity for mouse MBL-C due to the use of two monoclonal antibodies directed against different epitopes on the MBL-C molecule. - High sensitivity. The minimum concentration which can be measured is 0.82 ng/ml. - Large measurable concentration range. Standard curve from 0.82 - 200 ng/ml. - Efficient format:2 plates with twelve disposable 8-well strips allow free choice of batch size for the assay. - Standardization. The calibration standard has been standardized by Hbt. - Simple, rapid procedure. Four pipetting steps are required to complete the assay. Working time 3Â½ hours. For research purposes only. Caution: Not for use in humans.