LPCAT3 ELISA Kit (Mouse)

Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level Lysophosphatidylcholine Acyltransferase 3 (LPCAT3) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level Lysophosphatidylcholine Acyltransferase 3 (LPCAT3) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%

1-acylglycerophosphocholine O-acyltransferase;1-acylglycerophosphoethanolamine O-acyltransferase;1-acylglycerophosphoserine O-acyltransferase; C3f; Grcc3; Grcc3f; LPC acyltransferase 3; Lpcat; Lpeat; Lplat5; Lpsat; lyso-PC acyltransferase 3; lyso-PE acyltransferase; Lysophosphatidylcholine acyltransferase; Lysophosphatidylcholine acyltransferase 3; lysophosphatidylethanolamine acyltransferase; lysophosphatidylserine acyltransferase; lysophospholipid acyltransferase 5; lyso-PS acyltransferase; Mboa; Mboat5; Membrane-bound O-acyltransferase domain-containing protein 5; Moact5; Oac; Oact5; O-acyltransferase domain-containing protein 5; PTG.

Acyltransferase which mediates the conversion of lysophosphatidylcholine (1-acyl-sn-glycero-3-phosphocholine or LPC) into phosphatidylcholine (1,2-diacyl-sn-glycero-3-phosphocholine or PC) (LPCAT activity) . To a lesser extent, also catalyzes the acylation of lysophosphatidylethanolamine (1-acyl-sn-glycero-3-phosphoethanolamine or LPE) into phosphatidylethanolamine (1,2-diacyl-sn-glycero-3-phosphoethanolamine or PE) (LPEAT activity), and the conversion of lysophosphatidylserine (1-acyl-2-hydroxy-sn-glycero-3-phospho-L-serine or LPS) into phosphatidylserine (1,2-diacyl-sn-glycero-3-phospho-L-serine or PS) (LPSAT activity) . Favors polyunsaturated fatty acyl-CoAs as acyl donors compared to saturated fatty acyl-CoAs. Seems to be the major enzyme contributing to LPCAT activity in the liver. Lysophospholipid acyltransferases (LPLATs) catalyze the reacylation step of the phospholipid remodeling pathway also known as the Lands cycle.

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 Lysophosphatidylcholine Acyltransferase 3 (LPCAT3) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Lysophosphatidylcholine Acyltransferase 3 (LPCAT3) . 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 Lysophosphatidylcholine Acyltransferase 3 (LPCAT3), 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 Lysophosphatidylcholine Acyltransferase 3 (LPCAT3) 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) 89-104 96 EDTA Plasma (n=5) 91-103 99 Heparin Plasma (n=5) 78-94 87 Matrix Recovery range (%) Average (%) Matrix Recovery range (%) Average (%) Serum (n=5) 89-104 96 Serum (n=5) 89-104 96 EDTA Plasma (n=5) 91-103 99 EDTA Plasma (n=5) 91-103 99 Heparin Plasma (n=5) 78-94 87 Heparin Plasma (n=5) 78-94 87

Component Amount Anti-LPCAT3 Microplate 96 Wells (12 x 8 Well strips) LPCAT3 Lyophilized Standard 2 100X Biotinylated LPCAT3 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-LPCAT3 Microplate 96 Wells (12 x 8 Well strips) Anti-LPCAT3 Microplate 96 Wells (12 x 8 Well strips) LPCAT3 Lyophilized Standard 2 LPCAT3 Lyophilized Standard 2 100X Biotinylated LPCAT3 Detector Antibody 1 x 120 uL 100X Biotinylated LPCAT3 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 Lysophosphatidylcholine Acyltransferase 3 (LPCAT3) . No significant cross-reactivity or interference between Lysophosphatidylcholine Acyltransferase 3 (LPCAT3) and analogues was observed.