LIPA ELISA Kit (Mouse)

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

AA960673; acid cholesteryl ester hydrolase; cholesteryl esterase; L; Lal; Li; Lip1; Lip-1; lipase A; lysosomal acid lipase 1; lysosomal acid lipase/cholesteryl ester hydrolase; sterol esterase.

Crucial for the intracellular hydrolysis of cholesteryl esters and triglycerides that have been internalized via receptor-mediated endocytosis of lipoprotein particles. Important in mediating the effect of LDL (low density lipoprotein) uptake on suppression of hydroxymethylglutaryl-CoA reductase and activation of endogenous cellular cholesteryl ester formation (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 Lipase A, Lysosomal Acid (LIPA) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Lipase A, Lysosomal Acid (LIPA) . 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 Lipase A, Lysosomal Acid (LIPA), 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 Lipase A, Lysosomal Acid (LIPA) 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) 94-102 99 EDTA plasma (n=5) 88-103 94 heparin plasma (n=5) 80-93 86 Matrix Recovery Range (%) Average (%) Matrix Recovery Range (%) Average (%) serum (n=5) 94-102 99 serum (n=5) 94-102 99 EDTA plasma (n=5) 88-103 94 EDTA plasma (n=5) 88-103 94 heparin plasma (n=5) 80-93 86 heparin plasma (n=5) 80-93 86

Component Amount Anti-Lipa Microplate 96 Wells (12 x 8 Well Strips) Lipa Lyophilized Standard 2 x 20 ng 100X Biotinylated Lipa 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-Lipa Microplate 96 Wells (12 x 8 Well Strips) Anti-Lipa Microplate 96 Wells (12 x 8 Well Strips) Lipa Lyophilized Standard 2 x 20 ng Lipa Lyophilized Standard 2 x 20 ng 100X Biotinylated Lipa Detector Antibody 120 uL 100X Biotinylated Lipa 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 Lipase A, Lysosomal Acid (LIPA) . No significant cross-reactivity or interference between Lipase A, Lysosomal Acid (LIPA) and analogues was observed.