Mouse C3 (Complement Factor 3) Kit

Brand

GENWAY

Overview

Intended Use: The Complement factor 3 (C3) test kits are a highly sensitive two-site enzyme linked immunoassay (ELISA) for measuring C3 in serum or plasma of mice. Introduction: A number of serum proteins participate in acute inflammatory reactions. These include the complement coagulation and kinin systems as well as a number of other proteins known as acute phase proteins that regulate acute inflammation. The complement system is a complex set of up to 20 serum proteins. The most abundant and pivotal of the complement components is C3 which has a molecular weight of about 187 kD and consists of an alpha and beta chain. Principles of the assay: The principle of the double antibody sandwich ELISA is represented in Figure 1. In this assay the C3 present in samples reacts with the anti-C3 antibodies which have been adsorbed to the surface of polystyrene microtitre wells. After the removal of unbound proteins by washing anti-C3 antibodies conjugated with horseradish peroxidase (HRP) are added. These enzyme-labeled antibodies form complexes with the previously bound C3. Following another washing step the enzyme bound to the immunosorbent is assayed by the addition of a chromogenic substrate 3 3' 5 5'-tetramethylbenzidine (TMB). The quantity of bound enzyme varies directly with the concentration of C3 in the sample tested; thus the absorbance at 450 nm is a measure of the concentration of C3 in the test sample. The quantity of C3 in the test sample can be interpolated from the standard curve constructed from the standards and corrected for sample dilution. Storage and Stability: The expiration date for the package is stated on the box label. 1. DILUENTThe 5X Diluent Concentrate is stable until the expira-tion date. The 1X working solution is stable for at least one week from the date of preparation. Both solutions should be stored at 4-8 degree C. 2. WASH SOLUTIONThe 20X Wash Solution Concentrate is stable until the expiration date. The 1X working solution is stable for at least one week from the date of preparation. Both solutions can be stored at room temperature (16-25C) or at 4-8 degree C. 3. ENZYME-ANTIBODY CONJUGATEUndiluted horseradish peroxidase anti-C3 conjugate should be stored at 4-8C and diluted immediately prior to use. The working conjugate solution is stable for up to 8 hours. 4. CHROMOGEN-SUBSTRATE SOLUTIONThe Substrate Solution should be stored at 4-8 degree C and is stable until the expiration date. 5. STOP SOLUTIONThe Stop Solution should be stored at 4-8 degree C and is stable until the expiration date. 6. ANTI-C3 ELISA MICRO PLATEAnti-C3 coated wells are stable until the expiration date and should be stored at 4-8 degree C in sealed foil pouch with desiccant pack. 7. C3 CALIBRATORThe lyophilized C3 calibrator should be stored at 4 degree C or frozen until reconstituted. The reconstituted calibrator should be aliquoted out and stored frozen (Avoid multiple freeze-thaw cycles). The working standard solutions should be prepared immediately prior to use and are stable for up to 8 hours. INDICATIONS OF INSTABILITYIf the test is performing correctly the results observed with the standard solutions should be within 20 % of the expected values. Limitation of the Test: 1. Reliable and reproducible results will be obtained when the assay procedure is carried out with a complete understanding of the information contained in the package insert instructions and with adherence to good laboratory practice. 2. Factors that might affect the performance of the assay include proper instrument function cleanliness of glassware quality of distilled or deionized water and accuracy of reagent and sample pipettings washing technique incubation time or temperature. 3. Do not mix or substitute reagents with those from other lots or sources. Function: C3 plays a central role in the activation of the complement system. Its processing by C3 convertase is the central reaction in both classical and alternative complement pathways. After activation C3b can bind covalently via its reactive thioester to cell surface carbohydrates or immune aggregates. Function: Derived from proteolytic degradation of complement C3 C3a anaphylatoxin is a mediator of local inflammatory process. It induces the contraction of smooth muscle increases vascular permeability and causes histamine release from mast cells and basophilic leukocytes. Subunit: C3 precursor is first processed by the removal of 4 Arg residues forming two chains beta and alpha linked by a disulfide bond. C3 convertase activates C3 by cleaving the alpha chain releasing C3a anaphylatoxin and generating C3b (beta chain + alpha chain). During pregnancy C3dg exists as a complex (probably a 2:2:2 heterohexamer) with AGT and the proform of PRG2. Interacts with CR2 and VSIG4. Subcellular Location: Secreted. Ptm: C3b is rapidly split in two positions by factor I and a cofactor to form iC3b (inactivated C3b) and C3f which is released. Then iC3b is slowly cleaved (possibly by factor I) to form C3c (beta chain + alpha chain fragment 1 + alpha chain fragment 2) C3dg and C3f. Other proteases produce other fragments such as C3d or C3g. Polymorphism: There are two alleles: C3S (C3 slow) the most common allele in all races and C3F (C3 fast) relatively frequent in Caucasoids less common in Black Americans extremely rare in Orientals. Disease: Defects in C3 are the cause of C3 deficiency [MIM:120700]. It can result in susceptibility to pyogenic infection. Disease: Genetic variation in C3 is associated with susceptibility to age-related macular degeneration type 9 (ARMD9) [MIM:611378]. ARMD is a multifactorial eye disease and the most common cause of irreversible vision loss in the developed world. In most patients the disease is manifest as ophthalmoscopically visible yellowish accumulations of protein and lipid that lie beneath the retinal pigment epithelium and within an elastin-containing structure known as Bruch membrane. Similarity: Contains 1 anaphylatoxin-like domain. Similarity: Contains 1 NTR domain. Product Protocol: To see this product used in a peer reviewed journal click the following link:Opacity Proteins Increase Neisseria gonorrhoeae Fitness in the Female Genital Tract Due to a Factor under Ovarian Control

spezie

Mouse

detection target

C3 (Complement Factor 3)

pack size

1x96 Wells