For Western blot analysis, SDS–PAGE was carried out on AnykD (BioRad, Madrid, Spain) polyacrylamide gels, and 40 μg of protein extracts were transferred to nitrocellulose membranes using a semi-dry Trans-Blot cell (BioRad, Madrid, Spain). Membranes were used for cross-recognition assays using polyclonal antibodies anti-CrATG4 (Agrisera, Vännäs, Sweden) or anti-CrATG8A (Agrisera, Vännäs, Sweden). For immunodetection in membranes, a goat anti-rabbit IgG–horseradish peroxidase conjugate (Santa Cruz Biotechnology, Heidelberg, Germany) was used as the secondary antibody, together with an enhanced chemiluminescence kit (SupersignalTM West Picoplus, Fisher Scientific, Madrid, Spain). Densitometry of the different bands was performed using an image analyzer (Amersham Imager 600, GE Healthcare, Barcelona, Spain) and the Amersham ImageQuanTL 8.1 Program (Cytiva, Barcelona, Spain). Ponceau staining was used to correct the loading control in each line.
Goat anti rabbit igg horseradish peroxidase conjugate
Manufactured by Santa Cruz Biotechnology
Sourced in United States
Goat anti-rabbit IgG horseradish peroxidase conjugate is a secondary antibody that binds to rabbit primary antibodies. It is conjugated with the enzyme horseradish peroxidase, which can be used for signal amplification in various immunoassays and detection techniques.
Automatically generated - may contain errors
Lab products found in correlation
Amersham ecl select western blotting detection reagent, by GE Healthcare (3 mentions)
Dulbecco s modified eagle s medium dmem with l glutamine, by GenDEPOT (3 mentions)
Bca kit, by Thermo Fisher Scientific (3 mentions)
Celltiter 96 aqueous one solution cell proliferation assay kit, by Promega (2 mentions)
Hdac6, by Cell Signaling Technology (2 mentions)
Hdac6, by BPS Biosciences (2 mentions)
Hdac3, by BPS Biosciences (2 mentions)
Fatal bovine serum, by Thermo Fisher Scientific (2 mentions)
Trans blot cell, by Bio-Rad (1 mentions)
Anti cratg4, by Agrisera (1 mentions)
Supersignal west pico plus, by Thermo Fisher Scientific (1 mentions)
Anykd, by Bio-Rad (1 mentions)
Amersham imager 600, by GE Healthcare (1 mentions)
Nitrocellulose membrane, by Cytiva (1 mentions)
Glass bead, by Merck Group (1 mentions)
Bead beater, by Biospec (1 mentions)
Protease inhibitor cocktail, by Merck Group (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
Protein assay kit, by Bio-Rad (1 mentions)
X ray film, by Kodak (1 mentions)
8 protocols using goat anti rabbit igg horseradish peroxidase conjugate
1
Western Blot Analysis of Autophagy Proteins
2
Protein Extraction and Western Blot Analysis
3
Protein Expression Analysis in Liver Tissue
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Protein samples were prepared from liver tissues by homogenization with ice-cold lysate buffer for 1 h, and the tissue lysate samples centrifuged at 15,000 × g for 20 min at 4 °C. The supernatants were collected and the protein concentrations measured using Bradford assay (Bio-rad protein assay kit). For detection of SREBP-1c, SREBP-2, FAS, ACC, HMGCR and LXRα protein expressions, protein samples (60–80 μg) were denatured by boiling for 5 min, separated by 10% SDS-polyacrylamide gel, electroblotted at 4 °C and transferred onto polyvinylirdene diflouride (PVDF) membranes (Millipore, USA). The membranes were blocked in 5% non-fat milk for 2 h at room temperature and then incubated with rabbit polyclonal antibodies against SREBP-1c, SREBP-2, FAS, ACC, HMGCR and LXRα (diluted 1:1,000) with gentle agitation overnight at 4 °C. The membranes were washed 3 times for 10 min each with 15 mL of TBST and then incubated with second antibody (1:1000 goat Anti-rabbit IgG Horseradish Peroxidase Conjugate (Santacruz Biotechnology) at room temperature for 2 h. The protein was then visualized using an enhanced chemiluminescence solution and Kodak X-ray film. An imaging densitometer was used to scan the protein bands; quantification was performed using the Image Analysis Software.
4
Cellular Effects of Methamphetamine Exposure
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Dulbecco’s Modified Eagle’s medium (DMEM) with L-glutamine was purchased from GenDEPOT (Barker, TX, USA) and fatal bovine serum (FBS) and penicillin and streptomycin were purchased Gibco BRL (Gaithersburg, MD, USA). Antibodies for α-tubulin, Ac-α-tubulin (Lys40), Histone H3, Ac-Histone H3 (Lys9), and β-actin were purchased from Cell Signaling Technology (Boston, MA, USA). Goat anti-rabbit IgG horseradish peroxidase conjugate was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Cell Titer 96 Aqueous One Solution cell proliferation assay kit was purchased from Promega (Madison, WI, USA). Amersham ECL select Western blotting detection reagent was purchased from GE Healthcare. HDAC fluorogenic assay kits (HDAC1, #50061; HDAC6, #50076) were purchased from BPS Bioscience (San Diego, CA, USA). Tubastain A (Portland, OR, USA) was purchased from TCI chemicals. For in vitro studies, Methamphetamine (METH) was purchased from the Ministry of Food and Drug Safety (Cheongju, Korea).
5
HDAC Inhibitor Screening and Analysis
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Dulbecco’s Modified Eagle’s medium (DMEM) with L-glutamine was purchased from GenDEPOT (Barker, TX, USA) and RPMI 1640 medium, fatal bovine serum (FBS), and penicillin/streptomycin were purchased from Gibco BRL (Gaithersburg, MD, USA). Antibodies specific for α-tubulin, Ac-α-tubulin, Histone H3, Ac-Histone H3, PARP, caspase 3, cleaved caspase 8, β-actin HDAC1, and HDAC6 were purchased from Cell Signalling Technology (Boston, MA, USA). Rad52 antibody and goat anti-rabbit IgG horseradish peroxidase conjugate were purchased from Santa Cruz Biotechnology Inc. (Dallas, TX, USA). Cell Titre 96 Aqueous One Solution cell proliferation assay kit was purchased from Promega (Madison, WI, USA). Amersham ECL select Western blotting detection reagent was purchased from GE Healthcare (Waukesha, WI, USA). HDAC fluorogenic assay kits (HDAC1, HDAC3, HDAC6, and HDAC8) were purchased from BPS Bioscience (San Diego, CA, USA). OxiSelect™ Comet Assay Kit was purchased from Cell Biolabs, Inc (San Diego, CA, USA).
6
Vascular Oxidative Stress Measurement
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Vitamin D3 (1,25-dihydroxy vitamin D3, calcitriol) was purchased from Cayman Chemicals. 7-(1,3-Benzoxazol-2-ylsulfanyl)-3-benzyl-3H-[1,2,3]triazolo[4,5-d]pyrimidine (VAS2870), an inhibitor of NADPH oxidase, and Ang II (human) were from Sigma Aldrich. Material for the preparation of NO and ONOO− sensors: Mn (III) meso-tetra (N-methyl-4-pyridyl) porphyrin pentachloride, TMHPPMn (CAS # 125565–45-9) and nickel (II) tetrakis (3-methoxy-4-hydroxy-phenyl) porphyrin, TNMPPNi (Cat # T40113) (Frontier Scientific). MCDB-131 complete medium (VEC Technologies). Bovine serum albumin, primary antibodies: rabbit polyclonal immunoglobulin G (IgG) anti eNOS (SC-654), rabbit polyclonal IgG anti-inducible nitric oxide synthase (iNOS) (SC-650), rabbit polyclonal IgG anti-Nox4 (SC-30141) and goat anti-rabbit IgG–horseradish peroxidase conjugate (SC-2004, secondary antibody) (Santa Cruz bio-technologies). BCA kit (Thermo Fisher Scientific).
7
Radioactive Fatty Acid Metabolism
8
Histone Deacetylase Assays and Immunoblotting
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Dulbecco's modified Eagle's medium (DMEM) with L-glutamine was purchased from GenDEPOT (Barker, TX), fetal bovine serum was purchased from HyClone, and penicillin and streptomycin were purchased from Gibco BRL (Gaithersburg, MD). Antibodies for α-tubulin, Ac-α-tubulin (Lys40), Histone H3, Ac-Histone H3 (Lys9), HDAC1, HDAC6, and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) were purchased from Cell Signaling Technology (Boston, MA). Goat anti-rabbit IgG horseradish peroxidase conjugate was purchased from Santa Cruz Biotechnology (Santa Cruz, CA). Cell Titer 96 Aqueous One Solution cell proliferation assay kit was purchased from Promega (Madison, WI). Amersham ECL Select western blotting Detection Reagent was purchased from GE Healthcare. HDAC fluorogenic assay kits (HDAC1, #50061; HDAC2, #50062; HDAC3, #50073; HDAC4, #50064; HDAC6, #50076; HDAC11, #50687) were bought from BPS Bioscience (San Diego, CA).
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