DIO2 ELISA Kit (Rat)

Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level Deiodinase, Iodothyronine, Type II (DIO2) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level Deiodinase, Iodothyronine, Type II (DIO2) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%

5DII; deiodinase, iodothyronine, type 2; deiodinase, iodothyronine, type II; DIOII; type 2 DI; type II iodothyronine deiodinase; type-II 5'-deiodinase.

Catalyzes the deiodination of T4 (3,5,3',5'-tetraiodothyronine) into T3 (3,5,3'-triiodothyronine) . Essential for providing the brain with appropriate levels of T3 during the critical period of development.

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 Deiodinase, Iodothyronine, Type II (DIO2) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Deiodinase, Iodothyronine, Type II (DIO2) . 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 Deiodinase, Iodothyronine, Type II (DIO2), 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 Deiodinase, Iodothyronine, Type II (DIO2) 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) 95-102 99 EDTA Plasma (n=5) 78-102 90 Heparin Plasma (n=5) 80-94 86 Matrix Recovery range (%) Average (%) Matrix Recovery range (%) Average (%) Serum (n=5) 95-102 99 Serum (n=5) 95-102 99 EDTA Plasma (n=5) 78-102 90 EDTA Plasma (n=5) 78-102 90 Heparin Plasma (n=5) 80-94 86 Heparin Plasma (n=5) 80-94 86

Component Amount Anti-DIO2 Microplate 96 Wells (12 x 8 Well strips) DIO2 Lyophilized Standard 2 100X Biotinylated DIO2 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-DIO2 Microplate 96 Wells (12 x 8 Well strips) Anti-DIO2 Microplate 96 Wells (12 x 8 Well strips) DIO2 Lyophilized Standard 2 DIO2 Lyophilized Standard 2 100X Biotinylated DIO2 Detector Antibody 1 x 120 uL 100X Biotinylated DIO2 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 Deiodinase, Iodothyronine, Type II (DIO2) . No significant cross-reactivity or interference between Deiodinase, Iodothyronine, Type II (DIO2) and analogues was observed.