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MAp44 ELISA kit [Human]-2 x 96 det. (CAT#: CB-P082-K) Datasheet

Product Type
Kit
Description
The complement system is an important part of the innate immune response. It is composed of more than 30 proteins, which play a proteolytic cascade and are the first line of defense against pathogens. Complement also regulates B cell and T cell responses, thereby bridging innate immunity and adaptive immunity. Complement is activated in three ways, namely the classical pathway, the alternative pathway and the lectin pathway. The lectin pathway consists of a receptor that recognizes a variety of pathogens or apoptotic cells. The receptor mannan-binding lectin (MBL), H-ficolin, L-ficolin, M-ficolin or Collectin- K1 triggered. All molecules form complexes with MBL-related serine protease (MASP). So far, five MASP proteins have been identified: MASP1, MASP2, MASP3 and their truncated proteins MAp44 (also known as MAp1) and MAp19 (or sMAP). Both MASP1&3 and MAp44 are derived from the MASP1 gene. MAp44 mRNA consists of the first 8 exons, the first 8 exons encode the same first four domains as MASP-1 and MASP-3, and the other exon encodes 17 amino acids unique to MAp44. The lack of the lectin pathway is related to infection sensitivity, and the lectin-MASP complex is also involved in the activation of the coagulation system. The functions of MASP3, MAp44 and MAp19 in the lectin pathway are largely unknown. The competition between recombinant MAp44 and recombinant MBL or recombinant H-facolin complex led to a decrease in the cleavage level of C4 in serum depleted of MBL and H-facolin, respectively, indicating that MAp44 has an inhibitory effect on the activation of the complement system. MAp44 may represent a tissue-specific protection mechanism that can prevent local inflammation. The MBL-related protein MAp44 showed cardioprotection in the mouse model. Although related to cardiovascular risk factors, the level of MAp44 has nothing to do with the severity of human diseases. The concentration in human plasma is 0.8-3.2μg/ml. This assay uses a monoclonal antibody that recognizes the C-terminus to capture MAp44, and uses a detection antibody that reacts with the CCP-1 domain.
Applications
The human MAp44 ELISA kit is used for the in vitro quantitative determination of human MAp44 in serum and plasma samples.
Principle
The human MAp44 ELISA kit is a ready-to-use solid-phase enzyme-linked immunosorbent assay based on the sandwich principle. The working time is 3 hours. The effective form of the plate with twelve disposable 8-well plates allows free choice of batch size for determination. The samples and standards are incubated in microtiter wells coated with antibodies that recognize human MAp44. The biotinylated tracer antibody will bind to the captured human MAp44. 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. The standard curve was obtained by plotting the absorbance (linear) against the corresponding concentration (logarithm) of the human MAp44 standard. The concentration of the human MAp44 sample run at the same time as the standard can be determined from the standard curve.
Size
2 x 96 det.
Detection Method
Enzyme immunoassay (ELISA) technique
Detection Limit
1.6 to 100 ng/ml
Sensitivity
1.6 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
Cross Reactivity
Horse - Yes, Mouse - No, Pig - No, Rabbit - No, Rat - No
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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