PLA2G6 ELISA Kit (Rat)

Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level Phospholipase A2, Calcium Independent (iPLA2) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level Phospholipase A2, Calcium Independent (iPLA2) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%

2-lysophosphatidylcholine acylhydrolase;85 kDa calcium-independent phospholipase A2;85/88 kDa calcium-independent phospholipase A2; caI-PLA2; Group VI phospholipase A2; GVI PLA2; intracellular membrane-associated calcium-independent phospholipase A2 beta; iPla2; iPLA2-beta; palmitoyl-CoA hydrolase; patatin-like phospholipase domain-containing protein 9; phospholipase A2, group VI (cytosolic, calcium-independent) ; PNPLA9.

Catalyzes the release of fatty acids from phospholipids. It has been implicated in normal phospholipid remodeling, nitric oxide-induced or vasopressin-induced arachidonic acid release and in leukotriene and prostaglandin production. May participate in fas mediated apoptosis and in regulating transmembrane ion flux in glucose-stimulated B-cells. Has a role in cardiolipin (CL) deacylation. Required for both speed and directionality of monocyte MCP1/CCL2-induced chemotaxis through regulation of F-actin polymerization at the pseudopods (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 Phospholipase A2, Calcium Independent (iPLA2) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Phospholipase A2, Calcium Independent (iPLA2) . 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 Phospholipase A2, Calcium Independent (iPLA2), 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 Phospholipase A2, Calcium Independent (iPLA2) 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) 89-99 95 EDTA plasma (n=5) 99-105 102 heparin plasma (n=5) 78-94 84 Matrix Recovery Range (%) Average (%) Matrix Recovery Range (%) Average (%) serum (n=5) 89-99 95 serum (n=5) 89-99 95 EDTA plasma (n=5) 99-105 102 EDTA plasma (n=5) 99-105 102 heparin plasma (n=5) 78-94 84 heparin plasma (n=5) 78-94 84

Component Amount Anti-Pla2g6 Microplate 96 Wells (12 x 8 Well Strips) Pla2g6 Lyophilized Standard 2 x 20 ng 100X Biotinylated Pla2g6 Detector Antibody 120 uL Avidin/HRP Conjugate 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 TMB Substrate 1 x 9 mL Stop Solution 1 x 6 mL Component Amount Component Amount Anti-Pla2g6 Microplate 96 Wells (12 x 8 Well Strips) Anti-Pla2g6 Microplate 96 Wells (12 x 8 Well Strips) Pla2g6 Lyophilized Standard 2 x 20 ng Pla2g6 Lyophilized Standard 2 x 20 ng 100X Biotinylated Pla2g6 Detector Antibody 120 uL 100X Biotinylated Pla2g6 Detector Antibody 120 uL Avidin/HRP Conjugate 120 uL Avidin/HRP Conjugate 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 TMB Substrate 1 x 9 mL TMB Substrate 1 x 9 mL Stop Solution 1 x 6 mL Stop Solution 1 x 6 mL

This assay has high sensitivity and excellent specificity for detection of Phospholipase A2, Calcium Independent (iPLA2) . No significant cross-reactivity or interference between Phospholipase A2, Calcium Independent (iPLA2) and analogues was observed.