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C3b ELISA Kit [Mouse], 1 x 96 det. (CAT#: CB-P133-K) Datasheet

Product Type
Kit
Description
The complement system is an important part of innate immunity. Complement-derived products mediate the function of helping to kill and eliminate pathogens. However, inappropriate activation of the system can lead to the pathogenesis of immunological and inflammatory diseases. Complement component 3 (C3) occupies a central position due to the multiple biological activities of its activation fragment (including the main fragment C3b), which anchors the assembly of invertases that affect the activation of C3 and C5. C3 is converted to C3b through proteolysis of its anaphylatoxin domain by either of the two C3 convertases. This activates the stable thioester bond, causing C3b to covalently attach to the hydroxyl groups on the cell surface or protein surface through transesterification. C3b is further broken into iC3b, C3c, C3dg and C3f. C3b and iC3b act as opsonins, acting through different complement receptors, complement receptor 1 (CD35) and complement receptor 3 (CD11b / CD18).
The mouse C3b ELISA analysis is specific for cleaved C3 fragments C3b, iC3b and C3c and activated C3. Therefore, the positive reactivity in plasma or serum is related to the activation of the complement cascade and C3 lysis. In the event of an acute inflammatory response, a lot of C3 will be processed into products recognized by the assay, making the assay a useful tool for measuring acute inflammatory responses. This assay is less useful for assessing chronic inflammatory conditions, because minimal reactivity can be observed. In this case, it is mainly the C3dg product that stays in the inflammation site (C3c is cleared) that the analysis cannot identify. The long-term processing/activation of C3 is at a low level, which will reduce the detection of C3 fragments C3b, iC3b and C3c. Note that the level of complement activity depends on the mouse strain and may be affected by the way the sample is collected and processed. C3 levels may be different between healthy animals and diseased animals, and the optimal dilution between these states may also be different.
Applications
The Mouse C3b ELISA kit is used for the in vitro quantitative determination of C3b in serum and plasma samples.
Principle
Mouse C3b 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. Samples and standards are incubated in microtiter wells coated with antibodies that recognize mouse C3b. The biotinylated tracer antibody will bind to the captured Mouse C3b. 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 Mouse C3b standard. The mouse C3b sample concentration that runs simultaneously with the standard can be determined from the standard curve.
Size
1 x 96 det.
Detection Method
Enzyme immunoassay (ELISA) technique
Detection Limit
Measurable concentration range of 3.2 to 200 AU/ml.
Sensitivity
Minimum concentration which can be measured is 3.2 AU/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
Mouse
Cross Reactivity
Human - No, Pig - 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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