Biopulverizer

Manufactured by Biospec

Sourced in United States

The BioPulverizer is a laboratory equipment designed to homogenize and pulverize biological samples. It utilizes high-speed rotating blades to grind and disrupt the cellular structure of various types of samples, preparing them for further analysis or processing.

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Lab products found in correlation

Tissue tearor, by Biospec (8 mentions) Mini beadbeater 16, by Biospec (7 mentions) Agilent gene expression hybridization kit, by Agilent Technologies (4 mentions) Agilent microarray scanner, by Agilent Technologies (4 mentions) Trizol reagent, by Thermo Fisher Scientific (4 mentions) Pierce bca protein assay kit, by Thermo Fisher Scientific (4 mentions) Rneasy mini kit, by Qiagen (3 mentions) Hiseq 2500, by Illumina (3 mentions) Trizol, by Thermo Fisher Scientific (3 mentions) Bicinchoninic acid protein assay kit, by Thermo Fisher Scientific (2 mentions) Direct zol rna miniprep plus, by Zymo Research (2 mentions) Micro to midi total rna purification kit, by Thermo Fisher Scientific (2 mentions) Rna stat 60, by Tel-Test (2 mentions) Taqman reverse transcription reagent, by Thermo Fisher Scientific (2 mentions) Protease inhibitor cocktail, by Merck Group (2 mentions) Phosphate buffered saline (pbs), by Roche (2 mentions) Hanks balanced salt solution (hbss), by Thermo Fisher Scientific (2 mentions) Advanced protein assay, by Cytoskeleton (2 mentions) Complete protease inhibitor cocktail, by Roche (2 mentions) Gene expression hybridization kit, by Agilent Technologies (2 mentions)

71 protocols using biopulverizer

Quantitative Liver Lipid Profiling

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Liver wafers were pulverized into a fine powder by a stainless steel Bio-pulverizer (12 wells, capacity 10–100 mg per well, BioSpec Products, Bartlesville, OK) at the temperature of liquid N₂. Tissue fine powders (~ 20 mg) were weighed from each liver and homogenized in 10× diluted PBS by using 2.0-ml cryogenic vials (Corning Life Sciences, Tewksbury, MA). Protein assay on the homogenates was performed by using a bicinchoninic acid protein assay kit (Thermo Scientific, Rockford, IL) with bovine serum albumin as standards. All determined lipid levels were normalized to the protein content of individual samples.
Individual homogenate of the liver samples was accurately transferred into a disposable glass culture test tube. Internal standard (1,3-di15:0 DAG) for quantitation of DAG species was added prior to lipid extraction. This DAG species represents < 0.1% of the endogenous DAG mass levels as predetermined by ESI-MS. Lipid extraction was performed by using a modified Bligh and Dyer procedure as described²⁸. Each lipid extract was resuspended in a volume of 100 µl of CHCl₃/MeOH (1:1, v/v) per mg of tissue protein, flushed with N₂, capped, and stored at −20 °C.

Wang M., Hayakawa J., Yang K, & Han X. (2014). Characterization and Quantification of Diacylglycerol Species in Biological Extracts after One-step Derivatization: A Shotgun Lipidomics Approach. Analytical chemistry, 86(4), 2146-2155.

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Metabolite Extraction from Cells, Tissues, and Serum

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Cells were washed with ice-cold DPBS, followed by aspiration of medium. 500 μL of ice-cold extraction solvent (80% methanol and 20% water including 10 mM ammonium formate and 25 mM NEM, pH 7.0) was added to each well. After 30 min of incubation at 4°C, cells were scraped and, the supernatant moved to a 1.5mL tube and the debris cleared by centrifugation (17,000 g, 4°C, 20 min). Extracts were stored at −80°C until analysis. Cell numbers were counted by Scepter 2.0 cell counter and used to calculate intracellular metabolite concentrations. To extract metabolites from tissues, the frozen tissues were pulverized with a pre-chilled Bio-Pulverizer (59012MS, BioSpec). After weighing the tissues, the extraction solvent (80% methanol and 20% water including 10 mM ammonium formate and 25 mM NEM, pH 7.0) was added to the pulverized tissue for a final concentration of 50 mg tissue/mL extraction solvent for 30 min at 4°C. To extract metabolites from serum, 390 μL of extraction solvent (82% methanol and 18% water including 10 mM ammonium formate and 25 mM NEM, pH 7.0) was added to 10 μL of serum, followed by incubation at −80°C for 30 min. Debris was cleared by centrifugation (17,000 g, 4°C, 20 min). Extracts were stored at −80°C until analysis.

Born R.T, & Tootell R.B. (1991). Spatial frequency tuning of single units in macaque supragranular striate cortex.. Proceedings of the National Academy of Sciences of the United States of America, 88(16), 7066-7070.

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Muscle RNA Isolation and Purification

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Total muscle RNA was isolated from flash frozen samples using TRIzol/chloroform extraction after tissue homogenization with a biopulverizer (BioSpec Products, Inc, Fartlesville, Oklahoma) as previously described.22 Treatments with DNase were performed on columns (Direct‐zol RNA MiniPrep Plus, Zymo, Irvine, California) with DNase I (RNase‐free; New England BioLabs, Inc Ipswich, MA) according to manufacturer's instructions.

Valberg S.J., Soave K., Williams Z.J., Perumbakkam S., Schott M., Finno C.J., Petersen J.L., Fenger C., Autry J.M, & Thomas D.D. (2019). Coding sequences of sarcoplasmic reticulum calcium ATPase regulatory peptides and expression of calcium regulatory genes in recurrent exertional rhabdomyolysis. Journal of Veterinary Internal Medicine, 33(2), 933-941.

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Isolation and Analysis of Bone Gene Expression

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Both femur and tibia were isolated immediately after animals were euthanized. The epiphyses were then removed and bone morrow was flushed with PBS. The diaphyses were kept frozen in liquid nitrogen until processing. Frozen tissues were pulverized using a biopulverizer (Biospec Products, Inc., Bartlesville, OK, USA), followed by RNA extraction using RNA STAT60 (Tel-Test, Inc., Friendswood, TX, USA) and subsequent purification using Micro-to-Midi Total RNA Purification Kit (Invitrogen, Carlsbad, CA, USA). cDNA was synthesized using TaqMan Reverse Transcription Reagents (Applied Biosystems, Inc., Foster City, CA, USA) and random hexamer primers according to the recommendations of the manufacturer. Gene amplification was measured with SYBR Green using the ABI Prism ViiA^™ 7 real-time PCR System (Waltham, MA, USA). Analysis was carried out using the ViiA^™ 7 software supplied with the thermocycler. The sequences of the primer sets have been published previously [4 (link),23 (link),25 (link)]. The target gene expression was displayed normalized to GAPDH.

Wang L., Roth T., Abbott M., Ho L., Wattanachanya L, & Nissenson R.A. (2017). Osteoblast-derived FGF9 regulates skeletal homeostasis. Bone, 98, 18-25.

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Preparation of Human Amniotic Membrane Extracts

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Example 12

The entire procedure for preparation of total soluble human AM extracts (T) was carried out aseptically so as to be used for subsequent cell culture-based experiments. Frozen human placenta was obtained from Bio-Tissue, Inc. (Miami, Fla.), from which AM was retrieved. AM was sliced into small pieces to fit into the barrel of a BioPulverizer (Biospec Products, Inc., Bartlesville, Okla.), frozen in the liquid nitrogen, and then pulverized into a fine powder. The powder was weighed and mixed with cold PBS buffer (prepared by adding distilled H₂O to 1×PBS, pH7.4, from 10×PBS, cat #70011-044, Invitrogen, Carlsbad, Calif.) with protease inhibitors (protease inhibitor cocktail, P8340, Sigma, and supplemented with 1 mM PMSF) and phosphatase inhibitors (50 mM sodium fluoride and 0.2 mM sodium vanadate) at 1:1 (ml/g). The mixture was kept on ice and homogenized with a Tissue Tearor (Biospec Products, Inc., Dremel, Wis.) for 5 times, 1 min each with a 2 min interval cooling. This water-soluble extract was designated as “Total” (T). The total water-soluble extract was mixed for 1 hr at 4° C., centrifuged at 4° C. for 30 min at 48000×g. The supernatant was divided into aliquots and stored at −80° C.

US11318169B2. Compositions and methods for preventing the proliferation and epithelial-mesenchymal transition of epithelial cells (2022-05-03). TissueTech, Inc. [US]. Inventors: Scheffer Tseng [US], Ek Kia Tan [US], Hua He [US].

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Extraction of Total RNA from Frozen Tendon Tissue

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Frozen tendon tissues were homogenized in a Biopulverizer (Biospec, USA), which had been pre‐cooled in liquid nitrogen for 1 min. Frozen tendon tissue (10 mg) was placed into one of the designated little pits in the Biopulverizer and pulverized by blowing the hammer onto the pestle designed to match the pits. The tissue powder was transferred to an Eppendorf tube.
For 10 mg of tissue, 500 µL of PureZOL™ (Bio Rad, USA) was added to the tissue powder. The sample was vortexed for 5 min and incubated on ice for 30 min. It was then centrifuged for 10 min at 18 000 g at 4 °C. The supernatant was transferred to a new Eppendorf tube and 100 µL of chloroform (Sigma‐Aldrich, Austria) was added. The sample was mixed and incubated at room temperature (RT) for 5 min. The organic and aqueous phases were separated by high‐speed centrifugation (18 000 g at 4 °C for 15 min). The aqueous phase was collected into a new Eppendorf tube. Total RNA was recovered by the addition of isopropyl alcohol (Sigma, Austria) and glycerol (Thermo Scientific, USA). The mixture was incubated for 20 min on ice and centrifuged for 30 min at 18 000 g at 4 °C. Subsequently, the total RNA pellet was washed twice with 75% ETOH and solubilized in RNase‐free water. Digestion of genomic DNA was carried out with the Ambion™ DNA‐free™ DNA removal kit (Life Technologies, USA) for 30 min at 37 °C.

Ribitsch I., Gueltekin S., Keith M.F., Minichmair K., Peham C., Jenner F, & Egerbacher M. (2019). Age‐related changes of tendon fibril micro‐morphology and gene expression. Journal of Anatomy, 236(4), 688-700.

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Peanut Extract Preparation Protocol

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The peanut extract was prepared as previously described by Pi et al. (2015[22 (link)]). Raw peanuts were lightly blanched at 100 °C for 3 min and pulverized using a BioPulverizer (Biospec Products, Inc. Bartlesville, OK, USA). Pulverized peanuts were then mixed with phosphate buffered saline (Roche, Indianapolis, IN, USA) at a 1 kg:4 L ratio. A homogenizer (OMNI GLH) was used to further homogenize the peanut-PBS mixture. The mixture was stirred overnight at 4 °C, then centrifuged for 30 minutes at 100 x g at room temperature. The aqueous fraction was centrifuged for 30 more minutes at 100 x g at room temperature to remove residual lipids. The remaining aqueous fraction, or “peanut extract”, was collected and stored at -80 °C until use. A Pierce BCA Protein Assay Kit (Thermo Scientific, Rockford, IL, USA) was used to determine the protein concentration. The extract was stored so that each aliquot was thawed only once prior to use.

Beinart D., Ren D., Pi C., Poulton S., Holzknecht Z.E., Swanson C, & Parker W. (2017). Immunization enhances the natural antibody repertoire. EXCLI Journal, 16, 1018-1030.

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Protein Extraction from Human AM, Placenta, and Chorion

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Example 14

The complete procedure for preparation of protein extracts was carried out aseptically. Frozen human AM obtained from Bio-Tissue (Miami, Fla.) was briefly washed 2-3 times with HBSS (Invitrogen, Carlsbad, Calif.) to remove the storage medium. AM stroma was scraped from the stromal side of the AM by spatula for AM stroma extract preparation. Human placenta as well as chorion obtained from Baptist Hospital (Miami, Fla.) was rinsed 3 times with HBSS to remove blood. To prepare the water-soluble protein extract, total AM, scraped AM stroma, stroma-removed AM, placenta, and chorion were each frozen in the air phase of liquid nitrogen and each ground to fine particles using a BioPulverizer (Biospec Products, Inc., Bartlesville, Okla.) followed by homogenization on ice with Tissue Tearor (Biospec Products, Inc., Dremel, Wis.) in PBS (pH 7.4) for 1 min. Each homogenate was mixed for 1 h and centrifuged at 14,000 g for 30 min at 4° C. Each supernatant (in PBS) was then collected and stored in aliquots (0.5 ml) at −80° C. The BCA assay (Pierce, Rockford, Ill.) was used to quantitate the total protein in different extracts.

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Metabolic Profiling of Tissue Extracts

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After pulverization of liver and brain tissue with a pre-chilled BioPulverizer (59012MS, BioSpec), the extraction solvent (80% MeOH containing 2.49 μM ¹³C₃, ¹⁵n-serine, − 80 °C) was added for a final concentration of 50 mg tissue/mL and incubated for 24 hr at − 80 °C. The metabolite extracts were centrifuged (17,000 g, 4 °C, 20 min), and the supernatant was analyzed by LC-MS as previously described [18 ]. Data was acquired in ESI-positive mode. For targeted quantification of serine, ¹²c-serine and ¹³C₃, ¹⁵n-serine peak areas were manually integrated by EL-Maven (Version 0.6.1). Quantification was based on previous methods [4 (link)].

Kang Y.P., Falzone A., Liu M., González-Sánchez P., Choi B.H., Coloff J.L., Saller J.J., Karreth F.A, & DeNicola G.M. (2020). PHGDH supports liver ceramide synthesis and sustains lipid homeostasis. Cancer & Metabolism, 8, 6.

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Rapid Tissue Pulverization and DNA Extraction

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Frozen liver tissues were quickly pulverized with BioPulverizer (Bio Spec Products Inc.) on dry ice. DNA was extracted with Allprep DNA/RNA/Protein Mini Kit (Qiagen) according to the manufacturer’s protocol. DNA was further purified by phenol–chloroform extraction followed by ethanol precipitation.

Grimm S.A., Shimbo T., Takaku M., Thomas J.W., Auerbach S., Bennett B.D., Bucher J.R., Burkholder A.B., Day F., Du Y., Duncan C.G., French J.E., Foley J.F., Li J., Merrick B.A., Tice R.R., Wang T., Xu X., Bushel P.R., Fargo D.C., Mullikin J.C, & Wade P.A. (2019). DNA methylation in mice is influenced by genetics as well as sex and life experience. Nature Communications, 10, 305.

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