Srd5a2 ELISA Kit (Rat)

Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level Steroid 5 Alpha Reductase 2 (SRD5a2) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level Steroid 5 Alpha Reductase 2 (SRD5a2) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%

3-oxo-5-alpha-steroid 4-dehydrogenase 2;5 alpha-SR2; S5AR 2; SR type 2; Steroid 5-alpha-reductase 2; steroid-5-alpha-reductase, alpha polypeptide 2 (3-oxo-5 alpha-steroid delta 4-dehydrogenase alpha 2) .

Converts testosterone (T) into 5-alpha-dihydrotestosterone (DHT) and progesterone or corticosterone into their corresponding 5-alpha-3-oxosteroids (PubMed:1527072) . It plays a central role in sexual differentiation and androgen physiology (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 Steroid 5 Alpha Reductase 2 (SRD5a2) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Steroid 5 Alpha Reductase 2 (SRD5a2) . 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 Steroid 5 Alpha Reductase 2 (SRD5a2), 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 Steroid 5 Alpha Reductase 2 (SRD5a2) 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) 86-104 95 EDTA Plasma (n=5) 84-101 93 Heparin Plasma (n=5) 90-105 94 Matrix Recovery Range (%) Average (%) Matrix Recovery Range (%) Average (%) Serum (n=5) 86-104 95 Serum (n=5) 86-104 95 EDTA Plasma (n=5) 84-101 93 EDTA Plasma (n=5) 84-101 93 Heparin Plasma (n=5) 90-105 94 Heparin Plasma (n=5) 90-105 94

Component Amount Anti-Srd5a2 Microplate 96 Wells (12 x 8 Well Strips) Srd5a2 Lyophilized Standard 2 x 10 ng 100X Biotinylated Srd5a2 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-Srd5a2 Microplate 96 Wells (12 x 8 Well Strips) Anti-Srd5a2 Microplate 96 Wells (12 x 8 Well Strips) Srd5a2 Lyophilized Standard 2 x 10 ng Srd5a2 Lyophilized Standard 2 x 10 ng 100X Biotinylated Srd5a2 Detector Antibody 120 uL 100X Biotinylated Srd5a2 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 Steroid 5 Alpha Reductase 2 (SRD5a2) . No significant cross-reactivity or interference between Steroid 5 Alpha Reductase 2 (SRD5a2) and analogues was observed.