GPX4 ELISA Kit (Human)

Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level Glutathione Peroxidase 4 (GPX4) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level Glutathione Peroxidase 4 (GPX4) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%

Epididymis secretory sperm binding protein; Glutathione peroxidase 4; GPx-4; GSHPx-4; MCSP; PHGPx; phospholipid hydroperoxidase; phospholipid hydroperoxide glutathione peroxidase; phospholipid hydroperoxide glutathione peroxidase, mitochondrial; selenoprotein GPX4; SMDS; snGPx; snPHGPx; sperm nucleus glutathione peroxidase.

Essential antioxidant peroxidase that directly reduces phospholipid hydroperoxide even if they are incorporated in membranes and lipoproteins (By similarity) . Can also reduce fatty acid hydroperoxide, cholesterol hydroperoxide and thymine hydroperoxide (By similarity) . Plays a key role in protecting cells from oxidative damage by preventing membrane lipid peroxidation (By similarity) . Required to prevent cells from ferroptosis, a non-apoptotic cell death resulting from an iron-dependent accumulation of lipid reactive oxygen species (PubMed:24439385) . The presence of selenocysteine (Sec) versus Cys at the active site is essential for life: it provides resistance to overoxidation and prevents cells against ferroptosis (By similarity) . The presence of Sec at the active site is also essential for the survival of a specific type of parvalbumin-positive interneurons, thereby preventing against fatal epileptic seizures (By similarity) . May be required to protect cells from the toxicity of ingested lipid hydroperoxides (By similarity) . Required for normal sperm development and male fertility (By similarity) . Essential for maturation and survival of photoreceptor cells (By similarity) . Plays a role in a primary T-cell response to viral and parasitic infection by protecting T-cells from ferroptosis and by supporting T-cell expansion (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 Glutathione Peroxidase 4 (GPX4) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Glutathione Peroxidase 4 (GPX4) . 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 Glutathione Peroxidase 4 (GPX4), 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 Glutathione Peroxidase 4 (GPX4) 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) 82-94 91 EDTA Plasma (n=5) 83-92 87 Heparin Plasma (n=5) 82-92 85 Matrix Recovery range (%) Average (%) Matrix Recovery range (%) Average (%) Serum (n=5) 82-94 91 Serum (n=5) 82-94 91 EDTA Plasma (n=5) 83-92 87 EDTA Plasma (n=5) 83-92 87 Heparin Plasma (n=5) 82-92 85 Heparin Plasma (n=5) 82-92 85

Component Amount GPX4 Microplate 96 Wells (12 x 8 Well strips) GPX4 Lyophilized Standard 2 100X GPX4 HRP-Detector Anitbody 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 100X Luminol Substrate 1 x 2 mL Substrate Diluent 1 x 20 mL Component Amount Component Amount GPX4 Microplate 96 Wells (12 x 8 Well strips) GPX4 Microplate 96 Wells (12 x 8 Well strips) GPX4 Lyophilized Standard 2 GPX4 Lyophilized Standard 2 100X GPX4 HRP-Detector Anitbody 1 x 120 uL 100X GPX4 HRP-Detector Anitbody 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 100X Luminol Substrate 1 x 2 mL 100X Luminol Substrate 1 x 2 mL Substrate Diluent 1 x 20 mL Substrate Diluent 1 x 20 mL