PRDX6 ELISA Kit (Mouse)

Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level Peroxiredoxin 6 (PRDX6) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level Peroxiredoxin 6 (PRDX6) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%

1-cys;1-Cys peroxiredoxin;1-Cys Prx;1-cysPrx;9430088D19Rik; a; AA690119; acidic calcium-independent phospholipase A2; aiPLA2; antioxidant protein 2; anti-oxidant protein 2, related sequence 3; Aop2; Brp-; Brp-12; CP-; CP-3; glutathione-dependent peroxiredoxin; GP; GPx; LPC acyltransferase 5; LPCAT-5; Ltw; Ltw-; Ltw4; Ltw-4; Lvtw; Lvtw-4; lyso-PC acyltransferase 5; lysophosphatidylcholine acyltransferase 5; N; non-selenium glutathione peroxidase; NSGPx; ORF06; peroxiredoxin 5, related sequence 3; peroxiredoxin 6, related sequence 3; peroxiredoxin-6; Prdx5.

Thiol-specific peroxidase that catalyzes the reduction of hydrogen peroxide and organic hydroperoxides to water and alcohols, respectively. Can reduce H (2) O (2) and short chain organic, fatty acid, and phospholipid hydroperoxides. Also has phospholipase activity, and can therefore either reduce the oxidized sn-2 fatty acyl grup of phospholipids (peroxidase activity) or hydrolyze the sn-2 ester bond of phospholipids (phospholipase activity) . These activities are dependent on binding to phospholipids at acidic pH and to oxidized phospholipds at cytosolic pH. Plays a role in cell protection against oxidative stress by detoxifying peroxides and in phospholipid homeostasis.

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 Peroxiredoxin 6 (PRDX6) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Peroxiredoxin 6 (PRDX6) . 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 Peroxiredoxin 6 (PRDX6), 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 Peroxiredoxin 6 (PRDX6) 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) 78-94 86 EDTA Plasma (n=5) 88-98 94 Heparin Plasma (n=5) 89-101 92 Matrix Recovery range (%) Average (%) Matrix Recovery range (%) Average (%) Serum (n=5) 78-94 86 Serum (n=5) 78-94 86 EDTA Plasma (n=5) 88-98 94 EDTA Plasma (n=5) 88-98 94 Heparin Plasma (n=5) 89-101 92 Heparin Plasma (n=5) 89-101 92

Component Amount Anti-PRDX6 Microplate 96 Wells (12 x 8 Well strips) PRDX6 Lyophilized Standard 2 100X Biotinylated PRDX6 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-PRDX6 Microplate 96 Wells (12 x 8 Well strips) Anti-PRDX6 Microplate 96 Wells (12 x 8 Well strips) PRDX6 Lyophilized Standard 2 PRDX6 Lyophilized Standard 2 100X Biotinylated PRDX6 Detector Antibody 1 x 120 uL 100X Biotinylated PRDX6 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 Peroxiredoxin 6 (PRDX6) . No significant cross-reactivity or interference between Peroxiredoxin 6 (PRDX6) and analogues was observed.