PAOX ELISA Kit (Mouse)

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

2410012F02Rik; AI118225; exo-N4-amino; Pao; peroxisomal N (1) -acetyl-spermine/spermidine oxidase; peroxisomal N1-acetyl-spermine/spermidine oxidase; Polyamine oxidase.

Flavoenzyme which catalyzes the oxidation of N (1) -acetylspermine to spermidine and is thus involved in the polyamine back-conversion. Can also oxidize N (1) -acetylspermidine to putrescine. Substrate specificity: N (1) -acetylspermine = N (1) -acetylspermidine > N (1), N (12) -diacylspermine >> spermine. Does not oxidize spermidine. Plays an important role in the regulation of polyamine intracellular concentration and has the potential to act as a determinant of cellular sensitivity to the antitumor polyamine analogs.

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 Polyamine Oxidase (PAOX) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Polyamine Oxidase (PAOX) . 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 Polyamine Oxidase (PAOX), 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 Polyamine Oxidase (PAOX) 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) 84-101 89 EDTA plasma (n=5) 87-96 92 heparin plasma (n=5) 96-105 101 Matrix Recovery Range (%) Average (%) Matrix Recovery Range (%) Average (%) serum (n=5) 84-101 89 serum (n=5) 84-101 89 EDTA plasma (n=5) 87-96 92 EDTA plasma (n=5) 87-96 92 heparin plasma (n=5) 96-105 101 heparin plasma (n=5) 96-105 101

Component Amount Anti-Paox Microplate 96 Wells (12 x 8 Well Strips) Paox Lyophilized Standard 2 x 5 ng 100X Biotinylated Paox 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-Paox Microplate 96 Wells (12 x 8 Well Strips) Anti-Paox Microplate 96 Wells (12 x 8 Well Strips) Paox Lyophilized Standard 2 x 5 ng Paox Lyophilized Standard 2 x 5 ng 100X Biotinylated Paox Detector Antibody 120 uL 100X Biotinylated Paox 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 Polyamine Oxidase (PAOX) . No significant cross-reactivity or interference between Polyamine Oxidase (PAOX) and analogues was observed.