Common reagents, bovine serum albumin (BSA), streptavidin-agarose beads, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), and anti-HDAC1 antibody were purchased from Sigma-Aldrich (St. Louis, MO, USA). Cell culture supplies, fetal bovine serum (FBS), Lipofectamine 2000, and anti-SUMO1 antibody were purchased from Thermo Fisher Scientific (Waltham, MA, USA). Polyvinylidene difluoride (PVDF) membrane was purchased from Bio-Rad Laboratories (Hercules, CA, USA). Ultra ECL-HRP Substrate and the EasyPrep plasmid extraction kit were purchased from Biotools Co., Ltd. (Taipei, Taiwan). The Dual-Luciferase Reporter (DLR) Assay System was purchased from Promega (Madison, WI, USA). Anti-Sp1, anti-actin, anti-HDAC2, and anti-HDAC3 antibodies were purchased from Millipore (Billerica, MA, USA). Anti-acetyl-lysine, anti-SOD1, anti-SOD2, anti-catalase, anti-Prx3, and anti-GPx1 antibodies were purchased from Genetex Inc. (Irvine, CA, USA). Anti-p53, anti-phospho-p53 (Ser15), anti-acetyl-lysine, and anti-HDAC6 antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA). Anti-Nrf2 and horseradish peroxidase (HRP)-conjugated anti-mouse and HPR-conjugated anti-rabbit antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). The anti-Znf179 antibody was described previously [6] (link).
Anti gpx1
Manufactured by GeneTex
Sourced in United States
Anti-GPX1 is a primary antibody that recognizes the Glutathione Peroxidase 1 (GPX1) protein. GPX1 is an enzyme involved in the reduction of hydrogen peroxide and organic hydroperoxides, playing a role in cellular antioxidant defense mechanisms.
Automatically generated - may contain errors
Lab products found in correlation
Anti c caspase3, by Cell Signaling Technology (2 mentions)
Anti 4 hne, by Abcam (2 mentions)
Dual luciferase reporter dlr assay system, by Promega (1 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Anti actin, by Merck Group (1 mentions)
Streptavidin agarose beads, by Merck Group (1 mentions)
Anti sp1, by Merck Group (1 mentions)
Anti hdac2, by Merck Group (1 mentions)
Polyvinylidene difluoride pvdf membrane, by Bio-Rad (1 mentions)
Anti hdac6, by Cell Signaling Technology (1 mentions)
Anti sod1, by GeneTex (1 mentions)
Anti sod2, by GeneTex (1 mentions)
Anti catalase, by GeneTex (1 mentions)
Ecl kit, by Merck Group (1 mentions)
Anti β actin, by ZSGB-BIO (1 mentions)
Anti nf κb p65, by Cell Signaling Technology (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
Anti neun, by Cell Signaling Technology (1 mentions)
3 protocols using anti gpx1
1
Protein Extraction and Analysis Protocol
2
Protein Expression Analysis in Cultured SGNs
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After the drug treatment, the proteins from the cultured SGNs were extracted with radioimmunoprecipitation assay buffer (RIPA) buffer (Protein Biotechnology, China). The mixture was centrifuged at 4°C and 12,000 × g in a refrigerated centrifuge and the supernatant was collected. A total of 30 μg of each protein sample was denatured and separated by 12% sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) gels. The primary antibodies were anti-GPX1 (1:1,000 dilution, GeneTex, GTX116040), anti-C-caspase 3 (1:500 dilution, Cell Signaling Technology, 9664), anti-4-HNE (1:1000, Abcam, ab46545), anti-phosphorylated (p)-NFκB p65 (1:1,000 dilution, Cell Signaling Technology, 3033), and anti-β-actin (1:2,000 dilution, ZSGB-BIO, TA-09). The protein signals were detected using an ECL kit (Millipore, United States) and analyzed by Image J software. The relative optical density ratio was calculated by comparison to β-actin.
3
Immunohistochemical Analysis of Cochlear Tissues
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After organotypic culture or cryosection, the cochlear explants or tissue sections were fixed with 4% PFA, permeabilized with 1% TritonX-100 in PBS, and blocked by incubation in PBT-1 solution (0.1% Triton X-100, 8% donkey serum, 1% bovine serum albumin, and 0.02% sodium azide in PBS) at room temperature for 1 h. The samples were then incubated with the following primary antibodies: anti-Tuj 1 (1:1,000 dilution; Neuromics, MO15013), anti-GPX1 (1:500 dilution, GeneTex, GTX116040), anti-C-caspase 3 (1:400 dilution, Cell Signaling Technology, 9664), anti-NeuN (1:500, Cell Signaling Technology,12943), anti-4-HNE (1:500 dilution, Abcam, ab48506, United States), or anti-NF-κB p65 (1:500 dilution, Cell Signaling Technology, 6956) diluted in the blocking solution at 4°C overnight. The next day, samples were incubated with FITC-conjugated, TRITC-conjugated, or Cy5-conjugated (1:1000, Invitrogen, United States) secondary antibody along with 4′,6-diamidino-2-phenylindole (DAPI) (1:1000, Sigma-Aldrich, United States) at room temperature for 1 h. Then, coverslips were mounted and the samples were observed under a laser scanning confocal microscope (Leica SP8; Leica, Germany).
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