MBL ELISA Kit [Human], 1 x 96 det. (CAT#: CB-P113-K) Datasheet

Product Type
Kit
Description
Mannose-binding lectin (MBL) is a C-type lectin, an important part of innate immunity. MBL belongs to the collectin family of proteins and consists of a collagen-like domain and a carbohydrate recognition domain (CRD). In this way, MBL can recognize carbohydrates (such as mannose and N-acetylglucosamine) on pathogens. MBL forms oligomers of several sizes, which consist of three identical subunits of a 32 kDa polypeptide. MBL is a pattern recognition receptor. Once an infectious disease is recognized, MBL triggers the activation of the lectin complement pathway through attached C1r/C1s-like serine proteases (called MASP). The MBL-MASP complex proteolytically cleaves C4, C2 and C3. The lectin pathway is antibody- and C1q-dependent and therefore independent of the classical and alternative complement activation pathways. MBL is synthesized by hepatocytes and has been isolated from the liver or serum of several vertebrates. The concentration of MBL in normal human plasma ranges from 10 to 5,000 ng/ml. Up to 12% of healthy white blood donors have MBL concentrations below 100 ng/ml. Low plasma concentrations are associated with genetic defects in opsonization. The concentration of MBL increases in infectious diseases. In recurrent infections (especially in children), primary/secondary immunodeficiency, atherosclerosis/coronary heart disease, cystic fibrosis, transplantation, autoimmune diseases (SLE/rheumatoid arthritis) and habitual abortion, MBL can be detected. The determination of MBL is not affected by the mannan antibody. In human MBL analysis, any effect of the classical pathway is eliminated by using a special MBL binding buffer, which can inhibit the binding of C1q to immune complexes and destroy the C1 complex, while keeping the function of the MBL complex unchanged.
Applications
Human MBL (Lectin Assay) ELISA kit is used for in vitro quantitative determination of human MBL in serum, plasma and cell culture supernatant samples.
Principle
Human MBL ELISA kit is a ready-to-use solid-phase enzyme-linked immunosorbent assay based on the sandwich principle. The working time is 4 hours. The effective form of the plate with twelve disposable 8-well plates allows free choice of batch size for determination. After activation, the samples and standards will be captured by the consolidated mannan. The biotinylated tracer antibody will bind to the captured MBL. The streptavidin-peroxidase conjugate will bind the biotinylated tracer antibody. The streptavidin-peroxidase conjugate will react with the substrate tetramethylbenzidine (TMB). The enzyme reaction is stopped by adding oxalic acid. Measure the absorbance at 450nm with a spectrophotometer. Draw a standard curve by plotting the absorbance (linear) relative to the corresponding concentration (logarithm) of the MBL standard. The MBL concentration of samples run at the same time as the standard can be determined from the standard curve.
Size
1 x 96 det.
Detection Method
Enzyme immunoassay (ELISA) technique
Detection Limit
0.41-100 ng/ml
Sensitivity
0.41 ng/ml
Sample
Serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids
Specimen Collection
Blood samples were collected using sterile venipuncture techniques, and serum was obtained using standard procedures. At least 5 ml of whole blood is recommended. Let the blood clot in the serum test tube for 60-65 minutes at room temperature (20-25°C). Centrifuge the blood sample and transfer the cell-free serum to a clean test tube. The serum must be handled correctly to prevent complement activation in vitro. Serum should be frozen in a sealed tube at -70°C or lower for long-term storage or transport on dry ice. The sample should not be frozen and thawed more than once. Do not use jaundice, lipemia and hemolytic serum. You cannot use heat-inactivated serum. Plasma cannot be used.
Sample Volume
100 µl/well
Species Reactivity
Human
Protocol Length
2 hours
Storage
Product should be stored at 4 °C. Under recommended storage conditions, product is stable for at least six months.
Research Use
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.

All products and services are for Research Use Only. Do Not use in humans.

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