Rat Antithrombin III Kit

Brand

GENWAY

Overview

Intended Use: The ANTITHROMBIN III (AT3) test kits are a highly sensitive two-site enzyme linked immunoassay (ELISA) for measuring ANTITHROMBIN III in biological samples of ratsIntroduction: Antithrombin III is a small glycoprotein produced by the liver. It is a major inhibitor of the coagulation cascade and inhibits virtually all of the coagulation enzymes including Thrombin. Deficiency of Antithrombin III either genetic or acquired can lead to Thromboembolic disease. Principle of the test: The principle of the double antibody sandwich ELISA is represented in Figure 1. In this assay the ANTITHROMBIN III present in the sample reacts with the anti-ANTITHROMBIN III antibodies which have been adsorbed to the surface of polystyrene microtitre wells. After the removal of unbound proteins by washing anti-AT3 antibodies conjugated with horseradish peroxidase (HRP) are added. These enzyme-labeled antibodies form complexes with the previously bound serum AT3. 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 AT3 in the sample tested; thus the absorbance at 450 nm is a measure of the concentration of AT3 in the test sample. The quantity of AT3 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 expiration date. The 1X working solution is stable for at least one week from the date of preparation. Both solutions should be stored at 4-8C. 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 4-8C. 3. ENZYME-ANTIBODY CONJUGATEUndiluted horseradish peroxidase anti-AT3 conjugate should be stored at 4-8C and diluted immediately prior to use. The working conjugate solution is stable for one day. 4. CHROMOGEN-SUBSTRATE SOLUTIONThe Substrate Solution should be stored at 4-8C and is stable until the expiration date. 5. STOP SOLUTIONThe Stop Solution should be stored at 4-8C and is stable until the expiration date. 6. ANTI-RAT ANTITHROMBIN III ELISA MICRO PLATEAnti-Rat AT3 coated wells are stable until the expiration date and should be stored at 4-8C. 7. RAT ANTITHROMBIN III STANDARDSThe lyophilized Rat AntiThrombin III Calibrator should be stored at 4C or frozen until reconstituted. The reconstituted calibrator should be aliquoted and stored frozen (avoid multiple freeze-thaw cycles). The working standard solutions should be prepared immediately prior to use and are stable for 1 day. 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. Function: Most important serine protease inhibitor in plasma that regulates the blood coagulation cascade. AT-III inhibits thrombin as well as factors IXa Xa and XIa. Its inhibitory activity is greatly enhanced in the presence of heparin. Subcellular Location: Secreted extracellular space. Tissue Specificity: Found in plasma. Disease: Defects in SERPINC1 are the cause of antithrombin-III deficiency (AT3D) [MIM:107300]. AT3D is an important risk factor for hereditary thrombophilia a hemostatic disorder characterized by a tendency to recurrent thrombosis. AT3D is classified into 4 types. Type I: characterized by a 50% decrease in antigenic and functional levels. Type II: has defects affecting the thrombin-binding domain. Type III: alteration of the heparin-binding domain. Plasma AT-III antigen levels are normal in type II and III. Type IV: consists of miscellaneous group of unclassifiable mutations. Similarity: Belongs to the serpin family.

spezie

Rat

detection target

Antithrombin III

pack size

1x96 Wells