For Western blot (WB) analysis, DRG were homogenized in a lysis buffer (0.1 mol/L NaCl, 0.01 mol/L Tris-HCl, pH 7.5, 1 mmol/L EDTA, and 1 μg 7 Ml aprotinin) and the homogenates were centrifuged. Overall, 3 SOD1 and 3 control mice were used, respectively. The protein in the supernatants were subjected to SDS-PAGE electrophoresis and afterwards transferred to polyvinylidene difluoride membranes, stained with antibodies and visualized with the Odyssey system (LI-COR Biotechnology). Alpha-Tubulin was used as protein loading control. The antibodies used were rabbit polyclonal anti-SOD1 (dilution 1:1,000; Enzo, Life Sciences, Switzerland), goat polyclonal antibody anti-cathepsin D (dilution 1:2,000; Santa Cruz Biotechnology, USA) and mouse monoclonal anti-Tubulin (dilution 1:000; Abcam, Cambridge, USA). ImageJ software (U.S. National Institutes of Health, Bethesda, Maryland, USA) was used to quantify the density and size of the blots.
Anti sod1
Manufactured by Enzo Life Sciences
Sourced in Switzerland, United States, Germany
Anti-SOD1 is a laboratory product that detects the presence of Superoxide Dismutase 1 (SOD1) protein. SOD1 is an important antioxidant enzyme that helps protect cells from oxidative stress. The Anti-SOD1 product is designed to facilitate the identification and quantification of SOD1 levels in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Anti cathepsin d, by Santa Cruz Biotechnology (1 mentions)
Anti tubulin, by Abcam (1 mentions)
Odyssey system, by LI COR (1 mentions)
Rabbit anti gfap, by Agilent Technologies (1 mentions)
Alexa fluor 488 donkey anti rabbit, by Jackson ImmunoResearch (1 mentions)
Mouse anti gfap, by Novus Biologicals (1 mentions)
Anti gfap, by Novus Biologicals (1 mentions)
Anti mouse rage, by Merck Group (1 mentions)
Mouse anti neun, by Merck Group (1 mentions)
Anti rabbit and anti mouse igg peroxidase conjugated secondary antibodies, by Bio-Rad (1 mentions)
Anti rabbit iba 1, by Fujifilm (1 mentions)
Donkey anti mouse cy3, by Jackson ImmunoResearch (1 mentions)
To pro 3, by Thermo Fisher Scientific (1 mentions)
Anti gapdh, by Merck Group (1 mentions)
Donkey anti rabbit cy3, by Jackson ImmunoResearch (1 mentions)
Donkey anti mouse alexa fluor 488, by Jackson ImmunoResearch (1 mentions)
Anti mouse, by Jackson ImmunoResearch (1 mentions)
Anti erk1 2, by Santa Cruz Biotechnology (1 mentions)
Polyvinylidene fluoride pvdf membrane, by Merck Group (1 mentions)
Donkey anti rabbit, by Jackson ImmunoResearch (1 mentions)
4 protocols using anti sod1
1
Western Blot Analysis of SOD1 in DRG
2
Immunolabeling of Nervous System Markers
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The following primary antibodies were used for immunofluorescence: anti-rabbit S100B (1 : 7500, Novus Biological), anti-mouse S100B (1 : 1000, Sigma Aldrich), anti-mouse RAGE (1 : 200, Millipore), anti-rabbit GFAP (1 : 1000, Dako), anti-mouse GFAP (1 : 1000, Novus Biologicals), anti-mouse NeuN (1 : 500, Millipore), anti-rabbit Iba1 (1 : 200, Wako), anti-mouse CNPase (1 : 500, Novus Biologicals), and anti-rabbit ChAT (1 : 200, Millipore). Secondary fluorescent antibodies were Cy3 Donkey anti-rabbit (1 : 200), Alexa-Fluor 488 Donkey anti-rabbit (1 : 200), Cy3 Donkey anti-mouse (1 : 200), and Alexa Fluor 488 Donkey anti-mouse (1 : 200) from Jackson ImmunoResearch Laboratories. To-Pro-3 (1 : 10,000, Thermo Fisher Scientific) was used to stain nuclei. The primary antibodies used for Western blotting were anti-S100B (1 : 1000, Novus Biologicals), anti-rabbit RAGE (1 : 1000, Thermo Scientific), anti-mouse RAGE (1 : 1000, Millipore), anti-GAPDH (1 : 10,000, Millipore), anti-SOD1 (1 : 1000, Enzo Life Sciences), and anti-GFAP (1 : 5000, Novus biologicals). Anti-rabbit and anti-mouse IgG peroxidase-conjugated secondary antibodies (1 : 2500) were from Bio-Rad Laboratories.
3
Evaluation of Oxidative Stress Pathways
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QUIN, CUR, o-ophthaldehyde (OPA), NADPH, β-nicotinamide adenine dinucleotide phosphate (NADP+), GR, GSH, oxidized glutathione (GSSG), 2,3-naphthalenedicarboxyaldehyde (NDA), H2O2, 1-choloro-2,4-dinitrobenzene (CDNB), glucose 6-phosphate, dithiothreitol, bovine serum albumin (BSA), ethylenediamine tetraacetic acid (EDTA), paraformaldehyde (PAF), phenylmethylsulfonyl fluoride (PMSF), protease and phosphatase inhibitors, and primary antibody anti-α-tubulin were obtained from Sigma-Aldrich (St. Louis, MO, USA). Phosphoric acid (H3PO4) was obtained from Golden Bell Reagent (Guadalajara, Jalisco, Mexico). Fluoro-Jade B (FJ-B) and polyvinylidene fluoride (PVDF) membrane were obtained from Millipore (Bedford, MA, USA). Primary antibodies anti-Nrf2 (C-20), anti-GR, anti-γ-GCLc, anti-CAT, anti-phospho-ERK1/2, and anti-ERK1/2 were obtained from Santa Cruz Biotechnology Inc. (Dallas, TX, USA). Primary antibodies anti-BDNF and anti-GPx were obtained from Abcam (Cambridge, MA, USA). Primary antibodies anti-SOD1 and anti-SOD2 were obtained from Enzo Life Science (Farmingdale, NY, USA). Donkey anti-rabbit, anti-mouse, and anti-goat horseradish peroxidase-conjugate antibodies (secondary antibodies) were from Jackson Immunoresearch Laboratories Inc. (West Grove, PA, USA). Deionized water from a Milli-Q system (Millipore) was used for preparation of solutions.
4
Comprehensive Antibody Staining Protocol
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The following antibodies were used in this study: anti-FLAG (M2, Sigma Aldrich, St. Louis, MO), anti-HA (H6908, Sigma;3F10, Roche, Mannheim, Germany), anti-SOD1 (SOD100, Enzo Life Science, Farmingdale, NY; C17, Santa Cruz Biotechnology, Santa Cruz, CA), anti-chromogranin B (26102, QED Bioscience, San Diego, CA; PA1-10839, Thermo, Rockford, IL), B8H10 (33), C4F6 (37), anti-TGN38 (M290, Santa Cruz), anti-synaptophysin (D35E4, Cell signaling technology, Danvers, MA), anti- synaptotagmin V (46, Santa Cruz), anti-Akt1 (B1, Santa Cruz), anti-Mac2 (hybridoma from ATCC, Rockville, MD, USA), anti-glial fibrillary acidic protein (GFAP) (GA5, Cell signaling), anti-choline acetyltransferase (ChAT) (AB144P, Millipore, Billerica, MA), anti-neuronal nuclear antigen (NeuN) (MAB5294, Millipore), anti-actin (Millipore), anti-human protein gene product 9.5 (PGP9.5) (7863-0504, AbD Serotec, Raleigh, NC), anti-vesicular acetylcholine transport (VAChT) (Milipore), rhodamine conjugated α-Bungarotoxin (Invitrogen, Carlsbad, CA), anti-Bip (3177, Cell Signaling; ab21685, abcam, Cambridge, MA), anti-phospho-protein kinase-like endoplasmic reticulum kinase (PERK) (16F8, Cell Signaling), anti-C/EBP-homologous protein (CHOP) (F168, Santa Cruz), anti-caspase 12 (2202, Cell Signaling), SMI32 (Covance, Princeton, NJ), anti-SRY (E19, Santa Cruz).
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