DBI ELISA Kit (Mouse)

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

Binds medium- and long-chain acyl-CoA esters with very high affinity and may function as an intracellular carrier of acyl-CoA esters. It is also able to displace diazepam from the benzodiazepine (BZD) recognition site located on the GABA type A receptor. It is therefore possible that this protein also acts as a neuropeptide to modulate the action of the GABA receptor.

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 Diazepam Binding Inhibitor (DBI) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Diazepam Binding Inhibitor (DBI) . 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 Diazepam Binding Inhibitor (DBI), 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 Diazepam Binding Inhibitor (DBI) 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 (%) _x005F serum (n=5) 86-101 97 _x005F EDTA plasma (n=5) 90-104 97 _x005F heparin plasma (n=5) 81-99 87 _x005F Matrix Recovery Range (%) Average (%) Matrix Recovery Range (%) Average (%) serum (n=5) 86-101 97 serum (n=5) 86-101 97 EDTA plasma (n=5) 90-104 97 EDTA plasma (n=5) 90-104 97 heparin plasma (n=5) 81-99 87 heparin plasma (n=5) 81-99 87

Component Amount _x005F Anti-Dbi Microplate 96 Wells (12 x 8 Well Strips) _x005F Dbi Lyophilized Standard 2 x 10 ng _x005F 100X Biotinylated Dbi Detector Antibody 120 uL _x005F Avidin/HRP Conjugate 120 uL _x005F Standard Diluent 1 x 20 mL _x005F Detector Antibody Diluent 1 x 12 mL _x005F Conjugate Diluent 1 x 12 mL _x005F 30X Wash Buffer 1 x 20 mL _x005F TMB Substrate 1 x 9 mL _x005F Stop Solution 1 x 6 mL _x005F Component Amount Component Amount Anti-Dbi Microplate 96 Wells (12 x 8 Well Strips) Anti-Dbi Microplate 96 Wells (12 x 8 Well Strips) Dbi Lyophilized Standard 2 x 10 ng Dbi Lyophilized Standard 2 x 10 ng 100X Biotinylated Dbi Detector Antibody 120 uL 100X Biotinylated Dbi 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 Diazepam Binding Inhibitor (DBI) ._x005F No significant cross-reactivity or interference between Diazepam Binding Inhibitor (DBI) and analogues was observed.