Human HCC, normal liver samples, and cells were homogenized for protein extraction. Equal amount of the proteins was resolved on 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred to a polyvinylidene fluoride membrane. The membrane was blocked with 5% non-fat milk or 5% BSA for 1 hour and incubated with primary antibodies overnight at 4°C, followed by incubation with secondary antibodies for 1 hour at 37°C. The membrane was developed using the enhanced chemiluminescence method and imaged with a GEL DOC XR gel imager (Bio-Rad, Hercules, CA, USA). Using Quantity One software (Bio-Rad), the integrated density values of target protein bands were normalized by those of β-actin to obtain relative protein expression levels, except that the integrated density values of P-Akt bands were normalized by those of Akt bands to obtain the ratios of P-Akt/Akt.
Gel doc xr gel imager
Manufactured by Bio-Rad
Sourced in United States
The Gel Doc XR+ gel imager is a compact and versatile instrument designed for capturing high-quality images of agarose or polyacrylamide gels stained with fluorescent dyes. The system utilizes a sensitive CCD camera and advanced imaging software to produce detailed, reproducible images of DNA, RNA, or protein samples separated by electrophoresis.
Automatically generated - may contain errors
Lab products found in correlation
Molecular biology grade agarose, by Thermo Fisher Scientific (2 mentions)
Quantity one software, by Bio-Rad (1 mentions)
Eg1150h embedding machine, by Leica (1 mentions)
Polarstar plate reader, by BMG Labtech (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
E gel, by Thermo Fisher Scientific (1 mentions)
E gel electrophoresis system, by Thermo Fisher Scientific (1 mentions)
Accu chek blood glucose meter, by Roche (1 mentions)
Microplate reader, by Bio-Rad (1 mentions)
M365l2 c5, by Thorlabs (1 mentions)
9 protocols using gel doc xr gel imager
1
Western Blot Analysis of Protein Expression
2
Comprehensive Tissue Preparation and Analysis
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The following instruments were employed in the experiment: FreeZone-18 freeze dryer (Labconco Co., Kansas City, United States), ASP200S automatic tissue dehydrator (LEICA Co., Solms, Germany), BX53 intelligent biomicroscope (Olympus optical Co. Ltd., Tokyo, Japan), Leica RM2245 paraffin slicer (LEICA Co., Solms, Germany), GEL Doc XR Gel Imager (Bio-Rad Inc., California, United States), 85–2 magnetic mixer (Changzhou Guohua Instruments Co., Ltd., Changzhou, China), LDZX-50KBS high pressure steam sterilization pot (Shanghai Shenan Medical Device Factory, Shanghai, China), R-134A high-speed freeze centrifuge (Eppendorf Inc., Hamburg, Germany), EG1150H embedding machine (LEICA Co., Solms, Germany), Victor3V multifunction enzyme marker (PerkinElmer, Inc., San Francisco, California, United States), StepOnePlus real-time fluorescence quantitative PCR (Applied Biosystems Inc., California, United States), IMS-100 Ice Machine (Changshu Xueke Electric Appliances Co., Ltd., Changshu, China), and PowerPac Basic Gel Electrophoresis instrument (Bio-Rad Inc., California, United States).
3
Agarose Gel Electrophoresis of Nanoswitches
4
IC50 Determination of Ebselen and CuATSM
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IC50 of various drugs on untransfected NSC-34 cells was determined via a crystal violet assay (0.5 g/l crystal violet, 1% methanol [v/v], 1× PBS) as previously described (87 ). NSC-34 cells were treated for 48 h with ebslen and CuATSM at concentrations ranging from 0 to 500 μM with a final concentration of 1% (v/v) (dimethyl sulfoxide [DMSO]; Sigma–Aldrich). The drug-treated NSC34 cells were then fixed via the addition of prewarmed 4% paraformaldehyde in 1× PBS and incubated for 30 min at room temperature prior to the addition of crystal violet solution. Crystal violet–stained cells were imaged using a Gel Doc XR+ gel imager (Bio-Rad). Glacial acetic acid (100 μl 33% [v/v]) was used to release the crystal violet stain back into solution for quantification of absorbance at 590 nm on a POLARstar plate reader (BMG Labtech). The resulting data were plotted via Prism (GraphPad Prism, version 5.00 or version 8.00; GraphPad Software, Inc) using a log (inhibitor) versus normalized response variable slope fit.
5
Quantitative Nucleic Acid Analysis
6
Agarose Gel Electrophoresis of Nanoswitches
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Nanoswitches were run in 0.8% agarose gels, cast from molecular biology grade agarose (Fisher BioReagents) dissolved in 0.53× Tris-borate EDTA (TBE). Gels were typically run at 75 V (constant voltage) at room temperature. Samples were prestained by mixing 1× GelRed stain with the samples before loading. Gels were imaged with a Bio-Rad Gel Doc XR+ gel imager and analyzed using ImageJ.
7
Comprehensive Analysis Protocol
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The following equipment was used in this study: AB135-S standard analytical balance (Mettler-Toledo, Ltd., Columbus, OH, United States), ACCU-CHEK blood glucose meter (Roche, Basel, Switzerland), automatic biochemical analyzer (Rebenrily), microplate reader (Bio-RAD Laboratories, Hercules, CA, United States), real-time quantitative PCR (RT-qPCR) instrument (Thermo Fisher Scientific), Gel Doc XR gel imager (Bio-RAD Laboratories), Nano Drop 1000 nucleic acid quantifier (Thermo Fisher Scientific), vortex mixer (Qilin Bell Company, Jiangsu, China), and a tissue slicer (Thermo Fisher Scientific).
8
SDS-PAGE Analysis of O. oratoria MT-1
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SDS-PAGE was performed to check the purity of O. oratoria MT-1 and to determine its preliminary molecular weight (Mw). To prepare the sample, 100 μL of a 1.0 mg/mL O. oratoria MT-1 solution was heated in a water bath at 100 °C for 5 min and then centrifuged at 12,000 rpm for 5 min. Next, 10 µL of supernatant was used for SDS-PAGE electrophoresis analysis under the following conditions [18 (link)]: separation gel concentration, 12%; interlayer gel, 10%; spacer gel, 5%; and electrophoresis voltage, first 60 V and then 110 V after the sample had entered the separation gel; electrophoresis was stopped when the bromophenol blue band reached the bottom of the gel. Finally, the gel was recovered, stained for 1 h with 0.1-M Coomassie Brilliant Blue R-250 solution, and then scanned using a GelDoc XR gel imager (Bio-Rad, Hercules, CA, USA).
9
Optimizing DNA Irradiation Conditions
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A total of 50 ng of LA-DNA was irradiated with a 365 nm UV lamp (M365L2-C5 (Thorlabs) held 28.5 cm from the samples and collimated to a beam ~2.2 cm in diameter) for 0–15 min at 0.75 mW (measured at the center of the beam using a PM100A power meter console (Thorlabs) and S120VC photodiode sensor (Thorlabs)) and then combined with 6× loading dye and loaded into wells of a 1.5% TAE agarose gel. Gels were run in 1× TAE buffer at 100 V for 1 h and 15 min and then stained in 3× GelRed for up to 1 h. Gels were imaged using a BioRad Geldoc XR+ gel imager. The UV lamp setup described here was maintained in all remaining experiments.
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