Leukotriene A4 ELISA Kit

Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level Leukotriene A4 (LTA4) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level Leukotriene A4 (LTA4) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%

(2S- (2 alpha,3 beta (1E,3E,5Z,8Z) ) ) -3- (1,3,5,8-Tetradecatetraenyl) oxiranebutanoic acid. An unstable allylic epoxide, formed from the immediate precursor 5-HPETE via the stereospecific removal of a proton at C-10 and dehydration. Its biological actions are determined primarily by its metabolites, i.e., LEUKOTRIENE B4 and cysteinyl-leukotrienes. Alternatively, leukotriene A4 is converted into LEUKOTRIENE C4 by glutathione-S-transferase or into 5,6-di-HETE by the epoxide-hydrolase. (From Dictionary of Prostaglandins and Related Compounds, 1990)

This assay employs the competitive inhibition enzyme immunoassay technique. A monoclonal antibody specific to Leukotriene A4 (LTA4) has been pre-coated onto a microplate. A competitive inhibition reaction is launched between biotin labeled Leukotriene A4 (LTA4) and unlabeled Leukotriene A4 (LTA4) (Standards or samples) with the pre-coated antibody specific to Leukotriene A4 (LTA4) . After incubation the unbound conjugate is washed off. Next, avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. The amount of bound HRP conjugate is reverse proportional to the concentration of Leukotriene A4 (LTA4) in the sample. After addition of the substrate solution, the intensity of color developed is reverse proportional to the concentration of Leukotriene A4 (LTA4) in the sample.

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) 88-99 93 EDTA plasma (n=5) 95-104 101 heparin plasma (n=5) 81-88 85 Matrix Recovery Range (%) Average (%) Matrix Recovery Range (%) Average (%) serum (n=5) 88-99 93 serum (n=5) 88-99 93 EDTA plasma (n=5) 95-104 101 EDTA plasma (n=5) 95-104 101 heparin plasma (n=5) 81-88 85 heparin plasma (n=5) 81-88 85

Component Amount Anti-Leukotriene A4 Microplate 96 Wells (12 x 8 Well Strips) Leukotriene A4 Lyophilized Standard 2 100X Leukotriene A4 Biotin Complex 120 uL 100X Avidin/HRP Conjugate 120 uL Standard Diluent 1 x 20 mL Biotin Complex Diluent 1 x 12 mL Avidin/HRP Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL TMB Substrate 1 x 9 mL Stop Solution 1 x 6 mL Component Amount Component Amount Anti-Leukotriene A4 Microplate 96 Wells (12 x 8 Well Strips) Anti-Leukotriene A4 Microplate 96 Wells (12 x 8 Well Strips) Leukotriene A4 Lyophilized Standard 2 Leukotriene A4 Lyophilized Standard 2 100X Leukotriene A4 Biotin Complex 120 uL 100X Leukotriene A4 Biotin Complex 120 uL 100X Avidin/HRP Conjugate 120 uL 100X Avidin/HRP Conjugate 120 uL Standard Diluent 1 x 20 mL Standard Diluent 1 x 20 mL Biotin Complex Diluent 1 x 12 mL Biotin Complex Diluent 1 x 12 mL Avidin/HRP Conjugate Diluent 1 x 12 mL Avidin/HRP Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL 30X Wash Buffer 1 x 20 mL TMB Substrate 1 x 9 mL TMB Substrate 1 x 9 mL Stop Solution 1 x 6 mL Stop Solution 1 x 6 mL