P53 and α-tubulin antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA). ACSL4, SLC7A11, and NOX2 antibodies were purchased from Abclonal (Wuhan, China). Horseradish peroxidase (HRP)-conjugated IgG (H + L) secondary antibodies and CellROX Green Reagent were purchased from Thermo Scientific (Waltham, MA, USA). The Gpx4 antibody was obtained from Abcam (Cambridge, MA, USA). The Cytotox 96 nonradioactive cytotoxicity assay kit was purchased from Promega (Fitchburg, USA). Erastin was purchased from MCE (New York, NY, USA). The Lipid Peroxidation (MDA) Assay Kit, and GSSG/GSH quantification kits were purchased from Beyotime (Shanghai, China). Hoechst 33342 and one-step RT-qPCR kits were purchased from Sangon Biotech (Shanghai, China). Ferrostatin-1 (Fer-1) was purchased from Solarbio Science & Technology (Beijing, China). Precast Gel was purchased from Tsingke Biotechnology (Beijing, China). ChamQ Universal SYBR qPCR master mix was purchased from Vazyme Biotech (Nanjing, China). FerroOrange was purchased from Dojindo Molecular Technology (Kyushu, Japan). Gentamicin and other chemicals were from Sangon Biotech (Shanghai, China). All drug concentrations are expressed as the final molar concentration in working buffer.
Gpx4 antibody
Manufactured by Abcam
Sourced in United States
The GPX4 antibody is a laboratory reagent used for the detection and analysis of the GPX4 protein. GPX4 is an enzyme that plays a crucial role in the antioxidant defense system by reducing lipid hydroperoxides to their corresponding alcohols. This antibody can be used in various immunoassay techniques to study the expression and localization of GPX4 in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Cytotox 96 non radioactive cytotoxicity assay kit, by Promega (1 mentions)
α tubulin antibody, by Cell Signaling Technology (1 mentions)
Cellrox green reagent, by Thermo Fisher Scientific (1 mentions)
Chamq universal sybr qpcr master mix, by Vazyme (1 mentions)
Ferrostatin 1 fer 1, by Solarbio (1 mentions)
Ferroorange, by Dojindo Laboratories (1 mentions)
Acsl4, by ABclonal (1 mentions)
Slc7a11, by ABclonal (1 mentions)
Gentamicin, by Sangon (1 mentions)
Hoechst 33342, by Sangon (1 mentions)
Nrf2 antibody, by Abcam (1 mentions)
Paraformaldehyde solution, by Beyotime (1 mentions)
4 hne antibody, by Abcam (1 mentions)
Enhanced endogenous peroxidase blocking buffer, by Beyotime (1 mentions)
Minute total protein extraction kit, by Invent Biotechnologies (1 mentions)
Protein assay kit, by Abcam (1 mentions)
Enhanced chemiluminescence, by Beyotime (1 mentions)
Bicinchoninic acid (bca), by Abcam (1 mentions)
Precast gel, by Bio-Rad (1 mentions)
Bca assay, by Bio-Rad (1 mentions)
5 protocols using gpx4 antibody
1
Ferroptosis Induction and Measurement
2
Immunohistochemical Analysis of Nrf2, 4-HNE, and GPX4
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Lung tissues were fixed in 4% paraformaldehyde solution (Beyotime, China) for 24 h and then dehydrated and embedded in paraffin following routine methods. The slides were rinsed with Bond Dewax Solution at 72 °C to deparaffinize the tissue slices. Then, the sections were treated with Enhanced Endogenous Peroxidase Blocking Buffer (Beyotime) for 5 min to block endogenous peroxidase activity before they were incubated with Nrf2 antibody (1:500 dilution, Abcam, USA), 4-HNE antibody (1:200 dilution, Abcam) or GPX4 antibody (1:500 dilution, Abcam) overnight, after which DAB was quickly added. The color rendering time was controlled for 5 min. Images viewed under an Olympus microscope were captured. The Nrf2 immunostaining results were scored by multiplying the percentage of positive cells (0, <10%; 1+, 10–25%; 2+, 25–50%; 3+, 50–75% or 4+, >75%) by the staining intensity (0, negative; 1+, weak; 2+, moderate; or 3+, strong), and the 4-HNE and GXP4 immunostaining results were scored as the integrated optical density (IOD)/area as detected by Image-Pro Plus.
3
Protein Extraction and Western Blot Analysis in KGN Cells and PCOS Mice
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Total protein of KGN cells and PCOS mice was extracted by using Minute™ total protein extraction kit (Invent, Eden Prairie, MN, USA), and the concentration of protein was detected by BCA (bicinchoninic acid) protein assay kit (ABCAm). The primary antibodies were SKP2 antibody (1:500, ABCAm, ab183039), NEDD4L antibody (1:500, ABCAm, ab168349), GPX4 antibody (1:500, ABCAm, ab134953), UB antibody, and anti-GAPDH (glyceraldehyde-3-phosphate dehydrogenase) (1:5000, ABCAm, ab9485). The membranes were washed with TBST (TBS with Tween-20) for three times after incubation with primary antibodies for 2 h. Thereafter, membranes were incubated with goat anti-rabbit IgG H&L (HRP) (1:5000, ab205718) for 1 h at 37°C. The signals were visualized using enhanced chemiluminescence (Beyotime, Shanghai, China).
4
Protein Extraction and Western Blot Analysis
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Protein lysates were prepared from cells in RIPA buffer (25 mM Tris/HCl pH 7.4, 150 mM NaCl, 1% nonyl-phenoxylpolyethoxylethanol, 1% sodium deoxycholate, 0.1% sodium dodecyl sulfate) in the presence of protease/phosphatase inhibitor cocktail (Thermo Scientific, Rockford, Illinois, USA). Homogenates were further treated by three freeze/thawed cycles in liquid nitrogen, clarified by centrifugation at 13.2 K rpm, and protein concentration was determined by BCA assay (Bio-Rad, Hercules, California, USA). For immunoblot analysis, 20-30 µg of lysates were resolved on 4–15% or 10% precast gels (Bio-rad) and transferred onto 0.45 µm PVDF membrane (Immobilon-P; Merck Millipore) and probed with the specified antibody overnight at 4 °C. Secondary antibodies, conjugated with Horseradish peroxidase (HRP) were incubated at room temperature for 2 h. Proteins were visualized using ECL Plus (Pierce) or Immune-Star Western Chemiluminescence kit (Bio-rad). The following antibodies were used: caspase-2 (clone 11B4) (In-house), GPX4 antibody (Abcam, Cambridge, Massachusetts, USA), caspase-3 (Cell Signalling Technology, Danvers, Massachusetts, USA), p53 (DO-1 and 1801) (In-house), LC3-B (Cell Signalling Technology, Danvers, Massachusetts, USA) and Vinculin (Abcam, Cambridge, Massachusetts, USA).
5
GPX4 Expression Analysis by Flow Cytometry
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Cells were fixed with fixative disruptor (eBioscience, Cambridge, UK), stained with 1 mg/mL of GPX4 antibody (Abcam, Cambridge, UK), and then incubated with Alexa Fluor 647-labeled anti-rabbit GPX4 antibody according to the manufacturer's instructions, and rabbit IgG monoclonal (ab172730; Abcam) was used as primary isotype control. Cell sampls were tested on beckmancytoflex flow cytometer (BDBiosciences). Next, GPX4 expression was measured by flowcytometric analysis using FlowJo 10.4.2 software (Ashland, OR, USA).
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