C3b ELISA Kit [Mouse], 2 x 96 det. (CAT#: CB-P132-K) Datasheet

Kit

The complement system is an important part of innate immunity. Complement-derived products mediate functions and contribute to the killing and elimination of pathogens. However, inappropriate activation of the system can lead to the onset of immune and inflammatory diseases. Complement component 3 (C3) occupies the central position, because its activated fragments (including the main fragment C3b) have a variety of biological activities, anchoring the assembly of invertases that affect the activation of C3 and C5. C3 is affected by the proteolysis of its anaphylactoxin domain One of the C3 convertases converts into C3b which activates a stable thioester bond, causing C3b to be covalently attached to the cell surface or protein surface hydroxyl group through transesterification. C3b is further cleaved into iC3b, C3c, C3dg and C3f. C3b and iC3b plays the role of opsonin 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 in healthy and diseased animals, and the optimal dilution between these states may also be different.

The Mouse C3b ELISA kit is used for the in vitro quantitative determination of C3b in serum and plasma samples.

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.