Steel bead

Manufactured by Qiagen

Sourced in Germany

Steel beads are small, spherical particles made of stainless steel. They are designed for use in various laboratory applications that require agitation or disruption of samples, such as cell lysis or homogenization. The core function of steel beads is to provide a physical means of disrupting the structure of samples during processing or preparation.

Automatically generated - may contain errors

Lab products found in correlation

Tissuelyser 2, by Qiagen (3 mentions) Tissuelyser lt, by Qiagen (3 mentions) Nanodrop 1000, by Thermo Fisher Scientific (2 mentions) Mirneasy kit, by Qiagen (1 mentions) Collection tube, by Sarstedt (1 mentions) Rtl buffer, by Qiagen (1 mentions) Rnalater, by Thermo Fisher Scientific (1 mentions) Allprep dna rna protein, by Qiagen (1 mentions) Allprep dna rna protein mini kit, by Qiagen (1 mentions) Rtp dna rna virus mini kit, by Stratec (1 mentions) Prism 9, by GraphPad (1 mentions) Prism 7, by GraphPad (1 mentions) Realstar wnv rt pcr kit 1, by Altona Diagnostics (1 mentions) Qiaamp viral rna mini kit, by Qiagen (1 mentions) Trans blot turbo, by Bio-Rad (1 mentions) Complete protease inhibitor cocktail, by Roche (1 mentions) Mini protean tgx, by Bio-Rad (1 mentions) Vacuum centrifuge, by Eppendorf (1 mentions) Optima lc ms grade, by Thermo Fisher Scientific (1 mentions) Fastprep 24 homogenizer, by MP Biomedicals (1 mentions)

12 protocols using steel bead

Cortical Kidney Total RNA Extraction

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Total RNA from cortical kidney tissue was obtained using the miRNeasy kit (Qiagen) following the manufacturer’s instructions. Tissue was lysed using 700 mL QIAzol and homogenized with steel beads (Qiagen) using Tissue Lyser II (Qiagen) for 1 minute.

Connor K.L., Teenan O., Cairns C., Banwell V., Thomas R.A., Rodor J., Finnie S., Pius R., Tannahill G.M., Sahni V., Savage C.O., Hughes J., Harrison E.M., Henderson R.B., Marson L.P., Conway B.R., Wigmore S.J, & Denby L. (2020). Identifying cell-enriched miRNAs in kidney injury and repair. JCI Insight, 5(24), e140399.

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Cytokine Quantification in Cecal Tissue

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The ceca of uninfected and infected (21-h) mice were removed, slit longitudinally, and washed three times in PBS to remove the luminal contents. Tissue was transferred into 2 ml U-bottom tubes containing 1 ml of PBS and steel beads (Qiagen). The samples were weighed and subsequently homogenized with a MixerMill 301 homogenizer (Retsch, Newtown, PA). The homogenates were centrifuged at 12,000 × g for 10 min at 4°C, and the collected supernatant was stored at −80°C for later analysis. IL-1β and IFN-γ were quantified using commercially available ELISA kits (MBL and R&D Systems) according to the manufacturer’s protocols.

Yang H., de Souza Santos M., Lee J., Law H.T., Chimalapati S., Verdu E.F., Orth K, & Vallance B.A. (2019). A Novel Mouse Model of Enteric Vibrio parahaemolyticus Infection Reveals that the Type III Secretion System 2 Effector VopC Plays a Key Role in Tissue Invasion and Gastroenteritis. mBio, 10(6), e02608-19.

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Quantification of Viral Loads in Tissues

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Viral quantification was conducted by qRT-PCR and by Focus Forming Assay (FFA) on serum, spleen, kidney, liver, sciatic nerve, and brain. Sera were collected after centrifugation (15,900 g for 15 min at 4 °C) of blood harvested by cardiac puncture into collection tubes (Sarstedt, #41.1500.005). Following mouse perfusion with PBS, tissues were harvested and stored either in tubes containing RNA Later (Invitrogen) for qRT-PCR or in pre-weighed tubes containing MEMα medium and steel beads (Qiagen, #69989) for FFA. Tissues were then homogenized in RTL buffer (Qiagen) + 1% beta mercaptoethanol for qRT-PCR and MEMα medium for FFA followed by clarification (centrifugation at 2,000 × g for 5 min).

Branche E., Simon A.Y., Sheets N., Kim K., Barker D., Nguyen A.V., Sahota H., Young M.P., Salgado R., Mamidi A., Viramontes K.M., Carnelley T., Qiu H., Elong Ngono A., Regla-Nava J.A., Susantono M.X., Valls Cuevas J.M., Kennedy K., Kodihalli S, & Shresta S. (2019). Human Polyclonal Antibodies Prevent Lethal Zika Virus Infection in Mice. Scientific Reports, 9, 9857.

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Comprehensive Biomolecular Extraction from Breast Tumor Tissues

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Breast tumor tissues (PTs and IBTRs) were processed using the AllPrep DNA/RNA/Protein (Qiagen) protocol. Tissue lysis was performed by re-suspending ~30 mg of sliced frozen tissue in a solution containing 1% β-mercaptoethanol in RLT buffer (supplemented with antifoam agent; ID 19088, Qiagen). Next, steel beads (ID 79656, Qiagen) were added and samples were incubated in a Tissue Lyser LT (Qiagen) for 4 min at 50 Hz. steel beads were then removed and 400 µL of 1% β-mercaptoethanol in RLT buffer was added to samples, which were then centrifuged at 14,000 × g for 5 min. Supernatants were transferred to new tubes, and then frozen at −80 °C. DNA, RNA, and protein extraction were performed according to manufacturer instructions (AllPrep DNA/RNA/Protein minikit; Qiagen). Each spin column flowthrough (DNA, RNA, protein) was stored at −80 °C until analysis (sequencing or mass spectrometry; MS).

De Marchi T., Pyl P.T., Sjöström M., Reinsbach S.E., DiLorenzo S., Nystedt B., Tran L., Pekar G., Wärnberg F., Fredriksson I., Malmström P., Fernö M., Malmström L., Malmstöm J, & Niméus E. (2023). Proteogenomics decodes the evolution of human ipsilateral breast cancer. Communications Biology, 6, 139.

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Tick-Borne Virus RNA Extraction

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The extraction was carried out under the same conditions and in the same facility as used for human samples tested for CCHFV. Single whole ticks were placed in a 2 ml Eppendorf tube containing three to five steel beads (7 mm, Qiagen, Hilden, Germany), frozen on liquid nitrogen, and directly granulated in a Tissue-Lyser LT (Qiagen, Hilden, Germany) with a −20°C pre-cooled rotor at 50 Hz for 2–5 min. Repeated freezing and lysis was necessary in some cases. The samples were then re-suspended in 300–500 µl PBS (depending on tick size) containing 10% fetal calf serum, 500 IU/ml penicillin, and 500 µg/ml amphotericin, and centrifuged at 2,000 × g. For viral RNA extraction, 200 µl of the supernatant was diluted in 200 µl distilled water including carrier RNA and proteinase K (RTP DNA/RNA Virus Mini Kit, Stratec, Birkenfeld, Germany), following sample inactivation for 10 min in 95°C and DNA/RNA extraction according to the manufacturer’s instructions.

Sherifi K., Rexhepi A., Berxholi K., Mehmedi B., Gecaj R.M., Hoxha Z., Joachim A, & Duscher G.G. (2018). Crimean–Congo Hemorrhagic Fever Virus and Borrelia burgdorferi sensu lato in Ticks from Kosovo and Albania. Frontiers in Veterinary Science, 5, 38.

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Quantification of Organ Fungal Burden

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Mice were euthanized by CO₂ inhalation at the predetermined experimental endpoint. Organs from each animal were harvested and homogenized in sterile PBS using steel beads (Qiagen) and a bead beater. Organ homogenate was serially diluted in PBS. A 100-μL aliquot of each dilution was plated on antibiotic plates (YPD, 50 μg/mL ampicillin, and 30 μg/mL chloramphenicol), incubated for 48 to 72 h at 30°C, and assessed for CFU. CFU per gram of organ was plotted using GraphPad Prism 9. Statistical significance was calculated using an ordinary one-way analysis of variance (ANOVA) with Tukey’s multiple-comparison test.

Hoy M.J., Park E., Lee H., Lim W.Y., Cole D.C., DeBouver N.D., Bobay B.G., Pierce P.G., Fox D I.I.I., Ciofani M., Juvvadi P.R., Steinbach W., Hong J, & Heitman J. (2022). Structure-Guided Synthesis of FK506 and FK520 Analogs with Increased Selectivity Exhibit In Vivo Therapeutic Efficacy against Cryptococcus. mBio, 13(3), e01049-22.

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Quantification of Bacterial Colonization

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Luminal contents from the colon, cecum, and colon were placed in 2 ml U-bottom tubes containing 1 ml of sterile PBS and steel beads (Qiagen). All samples were weighed and subsequently homogenized with a MixerMill 301 homogenizer (Retsch, Newtown, PA). Six successive serial dilutions in PBS yielded 10⁻¹ to 10⁻⁶ dilutions. Ten microliters of each dilution was plated in triplicate onto MLB agar plates. Plates were incubated at 30°C overnight. The spleen, liver, and MLN were also collected from each animal and then homogenized and plated as outlined above. Plates were incubated at 30°C overnight.
Following incubation, bacteria were enumerated by counting the number of colonies per plate. The number of CFU per weight of the sample was calculated by multiplying the number of colonies by the dilution factor of the plate. Data were compiled in Microsoft Excel, and graphs were generated using Prism 7 (GraphPad).

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Viral RNA Extraction from Mosquito and Bird Organs

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RNA extraction from mosquito and bird organs: the mosquito pools (5-30 individuals) and 0.5 g of bird organ (0.1 g of brain, lung, heart, kidney, and lungs) were placed in a 2 ml Eppendorf tube containing three to ve steel beads (7 mm, Qiagen, Hilden, Germany), frozen on liquid nitrogen and directly pulverized in a Tissue-Lyser LT (Qiagen, Hilden, Germany) with a -20°C pre-cooled rotor at 50 Hz for 2-5 min. Repeated freezing and lysis was necessary in some cases. The samples were then re-suspended in 300-500 µl PBS (depending on tissue size) containing 500 IU/ml penicillin, 10% fetal calf serum, and 500 µg/ml amphotericin, and centrifuged at 2,000 × g. Viral RNA extraction was done with QIAamp ® Viral RNA Mini Kit (Qiagen, Hilden, Germany) according to the manufacturer's instructions.
RT-qPCR: 25 µl RNA of each sample extractions were tested using RealStar ® WNV RT-PCR Kit 1.0, (Altona Diagnostics, Hamburg, Germany) for the detection of West Nile virus (WNV) RNA, according to the manufacturer's instructions.

Rexhepi A., Sherifi K., Berxholi K., Xhekaj B., Muja-Bajraktari N., Özkul A., von Possel R, & Emmerich P. (2021). First Serological Evidence of West Nile Virus Among Equines and Birds in Kosovo, 2018-2019. Vector borne and zoonotic diseases (Larchmont, N.Y.), 21(2).

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Protein Extraction and Western Blotting

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For protein extraction, K562 cells and lung tissues were lysed with ice-cold RIPA buffer (ThermoFisher PI89901) supplemented with cOmplete Protease Inhibitor Cocktail (Roche). Lung tissues were homogenized using the Qiagen Tissue Lyser II (30 Hz, 1min) and steel beads (Qiagen) in RIPA buffer, followed by sonication. Equal amounts of protein (20–30µg) were mixed with 6X Laemmli SDS sample buffer (Fisher Scientific, AAJ61337AD). Samples were boiled at 95°C for 5min. Protein lysates were loaded on SDS-PAGE gels (Bio Rad Mini-PROTEAN^® TGX^™) and run at constant voltage. Proteins were transferred onto PVDF membranes using Trans-Blot Turbo (Bio Rad, 1704157) according to the manufacturer’s instructions. Membranes were blocked with 2% non-fat milk (Genesee Scientific Corporation 20–241) in TBST (Fisher Scientific 28360) for 1hr at RT on a rocker. The membranes were probed for primary antibodies overnight at 4°C. The corresponding secondary antibodies were applied to the membranes in 5% non-fat milk for 1hr at RT. The antibodies and concentrations are listed in Table S6. Bands were visualized using ECL (Fisher Scientific PI32106) according to the manufacturer’s instructions. All experiments were conducted in biological triplicates. One representative western blot is shown for each experiment.

Frandsen A, & Schousboe A. (1992). Mobilization of dantrolene-sensitive intracellular calcium pools is involved in the cytotoxicity induced by quisqualate and N-methyl-D-aspartate but not by 2-amino-3-(3-hydroxy-5-methylisoxazol-4-yl)propionate and kainate in cultured cerebral cortical neurons.. Proceedings of the National Academy of Sciences of the United States of America, 89(7), 2590-2594.

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Embryonic Metabolite Extraction Protocol

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Embryonic metabolism was quenched by adding 400μL cold 80:20 Methanol (Optima LC/MS Grade, Fisher Chemical):ddH2O to each mouse embryo on ice. Steel beads (Cat No. 69989, Qiagen) and a TissueLyser II (Qiagen) were used to lyse the embryos and extract metabolites. The homogenized samples were centrifuged at 14,000 rpm at 4°C for 10 min. The supernatant (i.e., the methanolic metabolite extract), was collected and the pellet was homogenized twice more in a fresh volume of 80:20 methanol:ddH₂O, followed by centrifugation. Supernatants from all successive homogenization and centrifugation steps was pooled for a total volume of 1.2 ml of metabolite extract, corresponding to each individual embryo. The extracts were dried using a vacuum centrifuge (Eppendorf) and the resulting metabolite films were stored at −80°C until LC/MS analysis. Mouse embryo tissue pellets were retained after centrifugation, dried and retained for protein quantification, as described below.

Akimova D., Wlodarczyk B.J., Lin Y., Ross M.E., Finnell R.H., Chen Q, & Gross S.S. (2017). Metabolite Profiling of Whole Murine Embryos Reveals Metabolic Perturbations Associated with Maternal Valproate-Induced Neural Tube Closure Defects. Birth defects research, 109(2), 106-119.

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