DGAT1 ELISA Kit (Human)

Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level Diacylglycerol-O-Acyltransferase Homolog 1 (DGAT1) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level Diacylglycerol-O-Acyltransferase Homolog 1 (DGAT1) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%

ACAT related gene product 1; ACAT-related gene product 1; acyl coenzyme A:cholesterol acyltransferase related gene 1; acyl-CoA retinol O-fatty-acyltransferase; acyl-CoA:diacylglycerol acyltransferase; ARAT; ARGP1; DGAT; diacylglycerol O-acyltransferase 1; DIAR7; diglyceride acyltransferase.

Catalyzes the terminal and only committed step in triacylglycerol synthesis by using diacylglycerol and fatty acyl CoA as substrates. In contrast to DGAT2 it is not essential for survival. May be involved in VLDL (very low density lipoprotein) assembly. In liver, plays a role in esterifying exogenous fatty acids to glycerol. Functions as the major acyl-CoA retinol acyltransferase (ARAT) in the skin, where it acts to maintain retinoid homeostasis and prevent retinoid toxicity leading to skin and hair disorders.

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 Diacylglycerol-O-Acyltransferase Homolog 1 (DGAT1) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Diacylglycerol-O-Acyltransferase Homolog 1 (DGAT1) . 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 Diacylglycerol-O-Acyltransferase Homolog 1 (DGAT1), 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 Diacylglycerol-O-Acyltransferase Homolog 1 (DGAT1) 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) 95-104 98 EDTA Plasma (n=5) 93-101 96 Heparin Plasma (n=5) 80-96 90 Matrix Recovery Range (%) Average (%) Matrix Recovery Range (%) Average (%) Serum (n=5) 95-104 98 Serum (n=5) 95-104 98 EDTA Plasma (n=5) 93-101 96 EDTA Plasma (n=5) 93-101 96 Heparin Plasma (n=5) 80-96 90 Heparin Plasma (n=5) 80-96 90

Component Amount Anti-DGAT1 Microplate 96 Wells (12 x 8 Well Strips) DGAT1 Lyophilized Standard 2 x 2 ng 100X Biotinylated DGAT1 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-DGAT1 Microplate 96 Wells (12 x 8 Well Strips) Anti-DGAT1 Microplate 96 Wells (12 x 8 Well Strips) DGAT1 Lyophilized Standard 2 x 2 ng DGAT1 Lyophilized Standard 2 x 2 ng 100X Biotinylated DGAT1 Detector Antibody 120 uL 100X Biotinylated DGAT1 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 Diacylglycerol-O-Acyltransferase Homolog 1 (DGAT1) . No significant cross-reactivity or interference between Diacylglycerol-O-Acyltransferase Homolog 1 (DGAT1) and analogues was observed.