Bead mill tube

Manufactured by Thermo Fisher Scientific

Sourced in United Kingdom

Bead mill tubes are laboratory equipment designed for the homogenization and disruption of biological samples. These tubes contain small beads that are used to physically break down samples, such as cells or tissues, through a process of mechanical agitation. The tubes are typically used in conjunction with a bead mill or homogenizer instrument to achieve effective sample preparation for various downstream analytical procedures.

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Lab products found in correlation

Trizol, by Thermo Fisher Scientific (2 mentions) Protein phosphatase inhibitor, by Abcam (1 mentions) Coomassie plus bradford assay reagent, by Thermo Fisher Scientific (1 mentions) Bio plex pro tgf β assay kit, by Bio-Rad (1 mentions) Pierce coomassie bradford assay kit, by Thermo Fisher Scientific (1 mentions) Bio plex 200, by Bio-Rad (1 mentions) Tissuelyser homogenizer, by Qiagen (1 mentions) Rneasy mini kit, by Qiagen (1 mentions)

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Bead mill

6 protocols using bead mill tube

Protein Extraction from Tissue Samples

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A total of 15 MT were added to RIPA lysis buffer supplemented with a complete protease inhibitor mixture and a protein phosphatase inhibitor (Abcam, UK). For tissue homogenisation, pre-filled bead mill tubes (Thermo Fisher, UK) and an evolution homogeniser was used (Percellys, France). The samples were thoroughly mixed and stored on ice for 10 min before centrifugation at 10,000 g for 10 min. The supernatants were transferred to a fresh tube and centrifuged for further 10 min period. The protein concentrations were measured utilising a Coomassie Plus Bradford assay reagent (Thermo Scientific, UK). The supernatants were stored at − 80 °C.

Kermanizadeh A., Valli J., Sanchez K., Hutter S., Pawlowska A., Whyte G., Moritz W, & Stone V. (2021). Particulate and drug-induced toxicity assessed in novel quadruple cell human primary hepatic disease models of steatosis and pre-fibrotic NASH. Archives of Toxicology, 96(1), 287-303.

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Quantifying TGFβ Ligand from Bone Lysates

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Protein from bone lysates (12-15 per age and sex) was harvested as previously described^{86 (link)} using Bead Mill Tubes (Thermo Fisher Scientific, Waltham, MA) containing radioimmunoprecipitation (RIPA) buffer with eComplete Mini protease inhibitor (Roche, Basil Switzerland). and homogenized using a benchtop bead mill homogenizer (Omni International, Kennesaw, GA) at 4 °C. Serum was collected from blood as previously described.^{27 (link)} Protein concentrations were quantified using the Pierce Coomassie (Bradford) Assay Kit (Thermo Fisher Scientific, Waltham, MA). Total levels of TGFβ ligand were quantified in hydrolyzed lysates by using the Bio-Plex Pro TGF-β Assay Kit (Bio-Rad, Hercules, CA) on a Bio-Plex 200 (Bio-Rad, Hercules, CA). TGFβ ligand levels were normalized to each sample’s original total protein concentration.

Schurman C.A., Kaya S., Dole N., Luna N.M., Castillo N., Potter R., Rose J.P., Bons J., King C.D., Burton J.B., Schilling B., Melov S., Tang S., Schaible E, & Alliston T. (2024). Aging impairs the osteocytic regulation of collagen integrity and bone quality. Bone Research, 12, 13.

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Quantifying Intracellular Energy Metabolites

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Relative amounts of creatine, phosphocreatine, ATP and ADP were measured using LC/MS from snap frozen tissue. 100ug tumor was homogenized in the tissue lyser homogenizer (Qiagen) in cold 2:1 LC/MS grade methanol:LC/MS grade water, in pre-filled bead mill tubes (Fisher scientific). Polar metabolites were isolated using cold LC/MS grade chloroform containing 50mM Metabolomics Amino Acid Standard (Cambridge Isotope Labs). The polar phase was collected, nitrogen – dried, and stored at −80°C until use. Targeted polar metabolomics was performed by The Rockefeller University Proteomics Resource Center.

Maguire O.A., Ackerman S.E., Szwed S.K., Maganti A.V., Marchildon F., Huang X., Kramer D.J., Rosas-Villegas A., Gelfer R.G., Turner L.E., Ceballos V., Hejazi A., Samborska B., Rahbani J.F., Dykstra C.B., Annis M.G., Luo J.D., Carroll T.S., Jiang C.S., Dannenberg A.J., Siegel P.M., Tersey S.A., Mirmira R.G., Kazak L, & Cohen P. (2021). Creatine mediated crosstalk between adipocytes and cancer cells regulates obesity-driven breast cancer. Cell metabolism, 33(3), 499-512.e6.

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Myometrial RNA Extraction Protocol

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Myometrial specimens were homogenized by the bead Mill 24 homogenizer (15–340-163, Fisher Scientific, Waltham, MA) in Bead Mill Tubes (15–340-154 Fisher Scientific, Waltham, MA) with 1 mL Trizol (15596026, Thermo Fisher Scientific, Waltham, MA). Tissue debris was pelleted and removed by centrifugation at 12000 ×g for 10 minutes at 4°C. After adding 200 uL 1-Bromo-3-chloropropane, samples were manually shake for 20 seconds followed by incubation at room temperature for 3 minutes. Phase separation was conducted by centrifugation at 12000 ×g for 18 minutes at 4°C. The RNA containing aqueous layer was retained and subsequently mixed with 500 uL 200 proof ethanol. The mixture was then passed through the RNA binding column of RNeasy Mini Kit (74104, Qiagen, Germantown, MD), followed by washing and elution steps described in the manufacture’s handbook.

Vostrov S.N., Kononenko O.V., Lubenko I.Y., Zinner S.H, & Firsov A.A. (2000). Comparative Pharmacodynamics of Gatifloxacin and Ciprofloxacin in an In Vitro Dynamic Model: Prediction of Equiefficient Doses and the Breakpoints of the Area under the Curve/MIC Ratio. Antimicrobial Agents and Chemotherapy, 44(4), 879-884.

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Tumor Extraction and Preservation

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Under isoflurane anesthesia, whole blood was collected by cardiac puncture and transferred to tubes containing 0.1 M citric acid (3:1 citric acid:blood) and stored at −20°C for LC-MS/MS analysis. Tumors were removed from mice flanks and cleared of surrounding mouse stroma. Tumor pieces between 50 mg and 100 mg were collected in a pre-weighed pre-filled bead mill tube (Fisher Scientific, Cat# 15-340-154) and then flash-frozen in liquid nitrogen. Other tumor fragments from vehicle- and compound-treated mice were placed in 10% neutral buffered formalin (NBF) within 2–3 minutes of surgical excision, fixed in NBF for 24 hours at room temperature and embedded in paraffin.

Xu H., George E., Gallo D., Medvedev S., Wang X., Kryczka R., Hyer M.L., Fourtounis J., Stocco R., Aguado-Fraile E., Petrone A., Yin S.Y., Shiwram A., Anderson M., Kim H., Liu F., Marshall C.G, & Simpkins F. (2024). Targeting CCNE1 amplified ovarian and endometrial cancers by combined inhibition of PKMYT1 and ATR. Research Square.

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Tumor Sampling and Preservation Protocol

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Under isoflurane anaesthesia, whole blood was collected by cardiac puncture and transferred to tubes containing 0.1 M citric acid (3:1 citric acid:blood) and stored at −20 °C for LC–MS/MS analysis. Tumours were removed from mice flanks and cleared of surrounding mouse stroma. Tumour pieces between 50 mg and 100 mg were collected in a pre-weighed pre-filled bead mill tube (Fisher Scientific, cat. no. 15-340-154) and then flash-frozen in liquid nitrogen. Other tumour fragments from vehicle- and compound-treated mice were placed in 10% neutral buffered formalin (NBF) within 2–5 min of surgical excision, fixed in NBF for 18–24 h at room temperature and embedded in paraffin.

Gallo D., Young J.T., Fourtounis J., Martino G., Álvarez-Quilón A., Bernier C., Duffy N.M., Papp R., Roulston A., Stocco R., Szychowski J., Veloso A., Alam H., Baruah P.S., Fortin A.B., Bowlan J., Chaudhary N., Desjardins J., Dietrich E., Fournier S., Fugère-Desjardins C., Goullet de Rugy T., Leclaire M.E., Liu B., Bhaskaran V., Mamane Y., Melo H., Nicolas O., Singhania A., Szilard R.K., Tkáč J., Yin S.Y., Morris S.J., Zinda M., Marshall C.G, & Durocher D. (2022). CCNE1 amplification is synthetic lethal with PKMYT1 kinase inhibition. Nature, 604(7907), 749-756.

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