BACE1 ELISA Kit (Rat)

Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level Beta-Site APP Cleaving Enzyme 1 (bACE1) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level Beta-Site APP Cleaving Enzyme 1 (bACE1) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%

Asp 2; ASP2; aspartyl protease 2; Bace; beta-secretase 1; beta-site amyloid precursor protein cleaving enzyme 1; beta-site APP cleaving enzyme 1; memapsin-2; membrane-associated aspartic protease 2.

Responsible for the proteolytic processing of the amyloid precursor protein (APP) . Cleaves at the N-terminus of the A-beta peptide sequence, between residues 671 and 672 of APP, leads to the generation and extracellular release of beta-cleaved soluble APP, and a corresponding cell-associated C-terminal fragment which is later released by gamma-secretase (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 Beta-Site APP Cleaving Enzyme 1 (bACE1) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Beta-Site APP Cleaving Enzyme 1 (bACE1) . 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 Beta-Site APP Cleaving Enzyme 1 (bACE1), 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 Beta-Site APP Cleaving Enzyme 1 (bACE1) 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) 85-92 89 EDTA Plasma (n=5) 78-99 92 Heparin Plasma (n=5) 80-97 93 Matrix Recovery range (%) Average (%) Matrix Recovery range (%) Average (%) Serum (n=5) 85-92 89 Serum (n=5) 85-92 89 EDTA Plasma (n=5) 78-99 92 EDTA Plasma (n=5) 78-99 92 Heparin Plasma (n=5) 80-97 93 Heparin Plasma (n=5) 80-97 93

Component Amount Anti-BACE1 Microplate 96 Wells (12 x 8 Well strips) BACE1 Lyophilized Standard 2 100X Biotinylated BACE1 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-BACE1 Microplate 96 Wells (12 x 8 Well strips) Anti-BACE1 Microplate 96 Wells (12 x 8 Well strips) BACE1 Lyophilized Standard 2 BACE1 Lyophilized Standard 2 100X Biotinylated BACE1 Detector Antibody 1 x 120 uL 100X Biotinylated BACE1 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 Beta-Site APP Cleaving Enzyme 1 (bACE1) . No significant cross-reactivity or interference between Beta-Site APP Cleaving Enzyme 1 (bACE1) and analogues was observed.