Whole-lung extracts were prepared from male mice lungs with nine volumes of ice-cold buffer (pH 7.4:5 mM phosphate buffer containing a complete protease inhibitor cocktail (Nacalai Tesque, Kyoto, Japan)) using Biomasher SP (Nippi Inc., Tokyo, Japan) for the thiobarbituric acid-reactive substances (TBA-RS) assay. Cytoplasmic and nuclear extracts were prepared using NE-PER™ nuclear and cytoplasmic extraction reagents (the protease and phosphatase inhibitors were added to CER I and NER) (Thermo Scientific, Rockford, IL, USA) using Biomasher II, according to the manufacturer’s instructions. Membrane extracts were prepared using the Mem-PER™ Plus Membrane Protein Extraction Kit (Thermo Scientific, Rockford, IL, USA) and Biomasher II according to the manufacturer’s instructions.
Biomasher 2
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Biomasher II is a versatile laboratory instrument designed for efficient tissue homogenization and sample preparation. It utilizes a motorized pestle to thoroughly disrupt various types of biological samples, including plant, animal, and microbial tissues. The Biomasher II is capable of generating homogenized samples suitable for downstream applications such as protein extraction, nucleic acid isolation, and enzymatic analysis.
Automatically generated - may contain errors
Lab products found in correlation
Ne per nuclear and cytoplasmic extraction reagent, by Thermo Fisher Scientific (2 mentions)
Trizol, by Thermo Fisher Scientific (2 mentions)
Complete protease inhibitor cocktail, by Nacalai Tesque (1 mentions)
Mem per plus membrane protein extraction kit, by Thermo Fisher Scientific (1 mentions)
Biomasher sp, by Nippi (1 mentions)
Biopulverizer, by Biospec (1 mentions)
Rneasy micro kit, by Qiagen (1 mentions)
2 protocols using biomasher 2
1
Lung Protein Extraction Techniques
2
Extraction and Purification of RNA from Rabbit Skin
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Skin samples (*50 mg/animal) from wild-type (n ¼ 4) and CS (n ¼ 4) rabbits were pulverized under liquid nitrogen using a BioPulverizer (BioSpec Products, Bartlesville, OK). Pulverized tissues were subsequently homogenized using a BioMasher II micro-homogenizer in 1 mL Trizol (Thermo Fisher Scientific, Grand Island, NY) reagent. Total RNA was precipitated according to the manufacturer's protocol. RNA precipitates were resuspended in 100 mL of Qiagen's RLT buffer and were column purified using the RNeasy Micro Kit (Qiagen, Valencia, CA) according to the manufacturer's protocol. Purified RNA was eluted in a final volume of 50 mL DEPC-treated dH 2 O.
First-strand cDNA synthesis was performed using the Accu-Script Hi-Fi cDNA Synthesis Kit (Agilent Technologies, Santa Clara, CA) according to the manufacturer's protocol. Firststrand cDNA synthesis reactions were performed using 1.5 to 5 mg purified RNA as template with an oligo(dT) primer. Incubations were performed for 2 hours at 42 C. Reactions were terminated by heat inactivation at 70 C for 15 minutes and samples were stored at -80 C until use.
First-strand cDNA synthesis was performed using the Accu-Script Hi-Fi cDNA Synthesis Kit (Agilent Technologies, Santa Clara, CA) according to the manufacturer's protocol. Firststrand cDNA synthesis reactions were performed using 1.5 to 5 mg purified RNA as template with an oligo(dT) primer. Incubations were performed for 2 hours at 42 C. Reactions were terminated by heat inactivation at 70 C for 15 minutes and samples were stored at -80 C until use.
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