GOT1 ELISA Kit (Bovine)

Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level Aspartate Aminotransferase (AST) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level Aspartate Aminotransferase (AST) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%

Aspartate aminotransferase 1; aspartate aminotransferase, cytoplasmic; cAspAT; cCAT; cysteine aminotransferase, cytoplasmic; cysteine transaminase, cytoplasmic; Glutamate oxaloacetate transaminase 1; glutamate oxaloacetate transaminase 1, soluble; transaminase A.

Biosynthesis of L-glutamate from L-aspartate or L-cysteine. Important regulator of levels of glutamate, the major excitatory neurotransmitter of the vertebrate central nervous system. Acts as a scavenger of glutamate in brain neuroprotection. The aspartate aminotransferase activity is involved in hepatic glucose synthesis during development and in adipocyte glyceroneogenesis. Using L-cysteine as substrate, regulates levels of mercaptopyruvate, an important source of hydrogen sulfide. Mercaptopyruvate is converted into H (2) S via the action of 3-mercaptopyruvate sulfurtransferase (3MST) . Hydrogen sulfide is an important synaptic modulator and neuroprotectant in the brain (By similarity) .

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Aspartate Aminotransferase (AST) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Aspartate Aminotransferase (AST) . Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Aspartate Aminotransferase (AST), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm +/- 10nm. The concentration of Aspartate Aminotransferase (AST) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Reconstitution & Storage Instructions Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Antibody Array (AA) Protocol

Matrix Recovery range (%) Average (%) Serum (n=5) 96-105 99 EDTA Plasma (n=5) 78-91 81 Heparin Plasma (n=5) 83-90 86 Matrix Recovery range (%) Average (%) Matrix Recovery range (%) Average (%) Serum (n=5) 96-105 99 Serum (n=5) 96-105 99 EDTA Plasma (n=5) 78-91 81 EDTA Plasma (n=5) 78-91 81 Heparin Plasma (n=5) 83-90 86 Heparin Plasma (n=5) 83-90 86

Component Amount Anti-GOT1 Microplate 96 Wells (12 x 8 Well strips) GOT1 Lyophilized Standard 2 100X Biotinylated GOT1 Detector Antibody 1 x 120 uL 100X Avidin-HRP Conjugate 1 x 120 uL Standard Diluent 1 x 20 mL Detector Antibody Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL Stop Solution 1 x 6 mL TMB Substrate 1 x 9 mL Component Amount Component Amount Anti-GOT1 Microplate 96 Wells (12 x 8 Well strips) Anti-GOT1 Microplate 96 Wells (12 x 8 Well strips) GOT1 Lyophilized Standard 2 GOT1 Lyophilized Standard 2 100X Biotinylated GOT1 Detector Antibody 1 x 120 uL 100X Biotinylated GOT1 Detector Antibody 1 x 120 uL 100X Avidin-HRP Conjugate 1 x 120 uL 100X Avidin-HRP Conjugate 1 x 120 uL Standard Diluent 1 x 20 mL Standard Diluent 1 x 20 mL Detector Antibody Diluent 1 x 12 mL Detector Antibody Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL 30X Wash Buffer 1 x 20 mL Stop Solution 1 x 6 mL Stop Solution 1 x 6 mL TMB Substrate 1 x 9 mL TMB Substrate 1 x 9 mL

This assay has high sensitivity and excellent specificity for detection of Aspartate Aminotransferase (AST) . No significant cross-reactivity or interference between Aspartate Aminotransferase (AST) and analogues was observed.