Avanti j 30i centrifuge

Manufactured by Beckman Coulter

Sourced in United States

The Avanti J-30I is a high-performance centrifuge designed for versatile laboratory applications. It features a powerful motor and a range of rotors to accommodate various sample volumes and tube sizes. The centrifuge provides excellent speed and force capabilities to facilitate efficient separation of samples.

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Lab products found in correlation

Lipofectamine 2000 transfection reagent, by Thermo Fisher Scientific (4 mentions) Ja 30.50 ti rotor, by Beckman Coulter (3 mentions) Lenti x concentrator, by Takara Bio (2 mentions) Beckman optima l 90k, by Beckman Coulter (1 mentions) Ti70 rotor, by Beckman Coulter (1 mentions) Pspax2, by Promega (1 mentions) Pmd2 g, by Promega (1 mentions) Polycarbonate tube, by Beckman Coulter (1 mentions) Vivaspin 500 column, by Sartorius (1 mentions) Pes syringe filter, by Merck Group (1 mentions) Phosphate buffered saline (pbs), by Merck Group (1 mentions) Vivaflow 200, by Sartorius (1 mentions) 6850 network gas chromatograph, by Agilent Technologies (1 mentions) Freeze dry system, by Labconco (1 mentions) Centrifuge 5810r, by Eppendorf (1 mentions) 0.2 m filter, by Cytiva (1 mentions) Ultra clear centrifuge tube, by Beckman Coulter (1 mentions) Padvantage, by Promega (1 mentions) Pspax2, by Addgene (1 mentions) Pmd2 g, by Addgene (1 mentions)

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Avanti J-30I (54 citations)

17 protocols using avanti j 30i centrifuge

Isolation of Grapefruit and Tomato Exosome-like Vesicles

Cited 3 times

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Fresh grapefruits and tomatoes were used as PEV sources. These fruits were purchased from a local market in Gatchina, Russia. The juices were extracted using a household juicer (Moulinex Y36-Vitafruit, Alençon, France), then each juice was filtered once using a PEV isolation technique, which was performed according to protocols in other studies [7 (link),15 (link),16 (link)].
Briefly, the filtered juices of grapefruits and tomatoes were sequentially centrifuged using an Avanti J30-I centrifuge (JA-10 rotor, Beckman Coulter, Brea, CA, USA) at 1500× g for 30 min, 3500× g for 20 min, 10,000× g for 60 min, 16,000× g for 60 min, and 10,000× g overnight to remove large particles and cellular debris. The supernatant was subjected to ultracentrifugation using a Beckman Optima L-90K ultracentrifuge (Ti 45 rotor, Beckman Coulter, Brea, CA, USA) at 150,000× g for 2 h. Then, the supernatant was removed, and the pellet was carefully resuspended in 2 mL of 1× PBS by gentle swaying overnight. The volume was adjusted to 10 mL with 1× PBS and ultracentrifuged at 150,000× g for 2 h (Ti 70 rotor, Beckman Coulter, Brea, CA, USA). The resulting pellet was resuspended with 1 mL of 1× PBS for at least 1 h at 4 °C. Final samples of grapefruit exosome-like vesicles (GEVs) and tomato exosome-like vesicles (TEVs) were aliquoted, snap frozen in liquid nitrogen, and stored at −80°C until analysis.

Kilasoniya A., Garaeva L., Shtam T., Spitsyna A., Putevich E., Moreno-Chamba B., Salazar-Bermeo J., Komarova E., Malek A., Valero M, & Saura D. (2023). Potential of Plant Exosome Vesicles from Grapefruit (Citrus × paradisi) and Tomato (Solanum lycopersicum) Juices as Functional Ingredients and Targeted Drug Delivery Vehicles. Antioxidants, 12(4), 943.

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Lentiviral Vector Production and Concentration

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HEK293T cells were grown in 10‐cm dishes to 80% confluency before transfection with the lentiviral vector (10 μg) with packaging vectors including pMD2.G (3 μg, Addgene plasmid # 12259), psPAX2 (6 μg, Addgene plasmid # 12260) and pAdVAntage (3 μg, E1711, Promega) using Lipofectamine 2000 Transfection Reagent (11668019, Thermo Fisher Scientific) according to the manufacturer's protocol. After 16 h, medium was refreshed. Supernatant containing lentivirus was harvested at 24 and 48 h after medium refreshing and pooled together. Supernatant was centrifuged at 300 g to remove cell fragments and passed through 0.45 μm filter. Lentivirus containing > 10 kb length insert (inducible CRISPRi) was concentrated using AVANTI J‐30I centrifuge (Beckman Coulter) with JS‐24.38 swing rotor at 72,000 g for 2 h at 4°C and pellets were dissolved in 200 μl PBS. Other lentivirus, including individual gRNAs, constructively active β‐catenin and SOX9 inducible overexpression, were concentrated using Lenti‐X™ Concentrator (631232, Takara) and pellets were dissolved in 400 μl PBS.

Sun D., Llora Batlle O., van den Ameele J., Thomas J.C., He P., Lim K., Tang W., Xu C., Meyer K.B., Teichmann S.A., Marioni J.C., Jackson S.P., Brand A.H, & Rawlins E.L. (2022). SOX9 maintains human foetal lung tip progenitor state by enhancing WNT and RTK signalling. The EMBO Journal, 41(21), e111338.

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Extracellular Vesicle Isolation Protocol

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Bacterial cells were removed from the culture broths by centrifugation twice at 7,000 × g for 10 min at 4°C followed by filtration through 0.22 μm PES syringe filter (Merck Millipore), and then concentrated using 100 kDa Vivaflow 200 (Sartorius AG). Crude input EV was prepared from the cleared supernatants by ultracentrifugation in polycarbonate tube (29 x 104 mm; Beckman Coulter) at 75,000 × g for 2.5 hr at 4°C in Avanti J-30I centrifuge with JA-30.50 Ti rotor (Beckman Coulter). Resulting EV pellets were resuspended in 1 ml of PBS (Sigma-Aldrich), filtered through 0.22 μm filter and further concentrated using 100 kDa Vivaspin 500 columns (Sartorius AG) before storage at −80°C [21]. These crude input EV preparations were used for further purification by either DGC or SEC as per [17]. Details are shown below and Supplementary Data 1.

Hong J., Dauros-Singorenko P., Whitcombe A., Payne L., Blenkiron C., Phillips A, & Swift S. (2019). Analysis of the Escherichia coli extracellular vesicle proteome identifies markers of purity and culture conditions. Journal of Extracellular Vesicles, 8(1), 1632099.

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Hydrogen Production Assay in Thermophilic Archaea

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Strains were grown in defined maltose media in 1 L culture bottles at 90°C with shaking. Cells were harvested by centrifugation at 18,000 × g for 10 min in a Beckman-Coulter Avanti J-30i centrifuge. Cell suspensions were created by washing harvested cells with an anaerobic resuspension buffer containing 20 mM imidazole, 30 mM MgCl₂·6H₂O, 0.5 M KCl, 2 mM cysteine-HCl, pH 6.5 and resuspending them in the same buffer at cell densities of OD₆₀₀ = 0.6. H₂ production assays are modified from that reported previously (Lim et al., 2014 (link)). Cell suspensions (final volume, 2 ml) were added to rubber-sealed glass 8 ml vials and the headspace was flushed with argon. Samples were incubated at 80°C for 3 min and the reaction was initiated by the addition of the desired concentration of NaCl from an anaerobic 2 M stock solution. At various time intervals, gas samples were taken and analyzed in a 6850 Network Gas Chromatograph (Agilent Technologies).

Haja D.K, & Adams M.W. (2021). pH Homeostasis and Sodium Ion Pumping by Multiple Resistance and pH Antiporters in Pyrococcus furiosus. Frontiers in Microbiology, 12, 712104.

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Curcumin-Loaded PLGA Nanoparticle Synthesis

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The PLGA based CUR nanoformulation was synthesized as described previously22 (link). Briefly, 90 mg of PLGA solution in 10 mL of acetone was mixed with 10 mg of CUR for 5 min. This solution was added dropwise to 20 mL of aqueous solution containing 1% wt/vol PVA and 10 mg of poly(l-lysine), over a period of 10 min on a magnetic stirrer at 800 rpm. The resulting suspension of Nano-CUR particles was stirred at room temperature for ~24 hrs to evaporate the acetone solvent completely. Larger aggregates and un-bound polymers were removed by centrifugation at 5000 rpm in an Eppendorf Centrifuge 5810 R (Eppendorf AG, Hamburg, Germany) for 10 min. Nano-CUR particles were recovered by ultracentrifugation at 30,000 rpm using a Rotor 30.50 in an Avanti J-30I Centrifuge (Beckman Coulter, Fullerton, CA, USA). Particles were washed twice and then freeze-dried using the Labconco Freeze Dry System (−48 °C, 133 × 10^–3 mBar; Labconco, Kansas City, MO). Control PLGA nanoparticles were similarly prepared by dissolving polymer in organic solvent without CUR, with the rest of the method being the same.

Zaman M.S., Chauhan N., Yallapu M.M., Gara R.K., Maher D.M., Kumari S., Sikander M., Khan S., Zafar N., Jaggi M, & Chauhan S.C. (2016). Curcumin Nanoformulation for Cervical Cancer Treatment. Scientific Reports, 6, 20051.

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Honey Bee Larval Fluid Preparation

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Larval bodily fluids were prepared by pooling third, fourth and fifth instars of the honey bee, Apis mellifera carnica, collected from the experimental beekeeping facility (Department of Physiology, Laboratory of Zoophysiology, University of Ghent, Belgium). Phenylthiourea (Hopkins & Williams Ltd) [100 µg per ml phosphate buffered saline (PBS, 20 mM KH₂PO₄, 60 mM Na₂HPO₄ and 145 mM NaCl, pH 7.2)], an inhibitor of phenoloxidase, was added immediately after squeezing the larvae to a final concentration of 10 µg/ml to prevent melanization. The homogenate was centrifuged twice for 30 min at 75,600×g and 4°C with an Avanti J30-I centrifuge (Beckman Coulter, Inc.). The clarified supernatant was filter sterilized with a 0.2 µm filter (Whatman) to render the bodily fluid for P. larvae culture spiking.

De Smet L., De Koker D., Hawley A.K., Foster L.J., De Vos P, & de Graaf D.C. (2014). Effect of Bodily Fluids from Honey Bee (Apis mellifera) Larvae on Growth and Genome-Wide Transcriptional Response of the Causal Agent of American Foulbrood Disease (Paenibacillus larvae). PLoS ONE, 9(2), e89175.

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Ultracentrifugation of Serum for EV Isolation

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A volume of 0.25 ml of human serum was diluted to 32 ml in phosphate-buffered saline (PBS) and transferred to a 35 ml open top Ultra-Clear^TM centrifuge tube (Beckman Coulter, Brea, CA) and centrifuged for 1 h at 100,000g at 4°C in a Beckman Coulter Avanti j-30 i centrifuge (Js-24,38 rotor, Beckman Coulter). The pellet was resuspended in 33 ml of PBS for a washing step and centrifuged again under the same conditions. Finally, the pellet was resuspended in H₂O (250 µl) for subsequent RNA extraction or EV analysis.

Andreu Z., Rivas E., Sanguino-Pascual A., Lamana A., Marazuela M., González-Alvaro I., Sánchez-Madrid F., de la Fuente H, & Yáñez-Mó M. (2016). Comparative analysis of EV isolation procedures for miRNAs detection in serum samples. Journal of Extracellular Vesicles, 5, 10.3402/jev.v5.31655.

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Bacterial Isolate Promotes Arabidopsis Growth

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The effect of the isolated bacterial strain on promoting plant growth was analyzed in Arabidopsis thaliana ecotype Columbia-0 (Col-0). Four-day-old Arabidopsis seedlings grown in the Murashige and Skoog (MS) medium were co-cultured with the bacterial inoculants (5 colonies of BcD1) for 7 days, and root development of the treated seedlings was analyzed, including the number of lateral roots and root hairs. The growth-promoting effect of the isolated bacterial strain was further analyzed in 2-week-old Arabidopsis seedlings grown in soil. The bacterial isolate cultured in medium containing 0.5% sucrose, 0.5% peptone, 0.5% MgSO₄, 0.04% KH₂PO₄, 0.03% K₂HPO₄, and 0.5% yeast extract at 28 °C in the dark for 24 h was centrifuged with the Avanti J-30I centrifuge (Beckman Coulter) at 5000× g for 10 min. The resulting bacterial pellet was resuspended in water to a density of 1 × 10⁸ CFU/mL and this bacterial suspension was used for plant treatment via foliar spray. Bacterial treatment was performed once a week, and the chlorophyll content and fresh weight of the seedlings were recorded after three treatments. Chlorophyll content was determined as described by Kurniawan et al. [42 (link)].

Tsai S.H., Hsiao Y.C., Chang P.E., Kuo C.E., Lai M.C, & Chuang H.W. (2023). Exploring the Biologically Active Metabolites Produced by Bacillus cereus for Plant Growth Promotion, Heat Stress Tolerance, and Resistance to Bacterial Soft Rot in Arabidopsis. Metabolites, 13(5), 676.

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Lentiviral Vector Production and Concentration

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HEK293T cells were kindly provided by Dr. Rick Livesey and tested mycoplasma negative. We grew HEK293T cells in 10 cm dishes to a confluence of 80% before we transfected the lentiviral vector (10 μg) with packaging vectors including pMD2.G (3 μg, Addgene plasmid # 12259), psPAX2 (6 μg, Addgene plasmid # 12260) and pAdVAntage (3 μg, E1711, Promega) using Lipofectamine 2000 Transfection Reagent (11668019, Thermo Fisher Scientific) according to the manufacturer’s protocol. After 16 hr, medium was refreshed. Supernatant containing lentivirus was harvested at 24 hr and 48 hr after medium refreshing and pooled together. Supernatant was centrifuged at 300 g to remove cell fragments and passed through 0.45 μm filter. Lentivirus containing >10 kb length insert (inducible CRISPRi and CRISPRa) was concentrated using AVANTI J-30I centrifuge (Beckman Coulter) with JS-24.38 swing rotor at 72,000 g for 2 hr at 4°C and pellets were dissolved in 200 μl PBS. Other lentivirus, including gRNA and NKX2-1-inducible overexpression, were concentrated using Lenti-X Concentrator (631232, Takara) and pellets were dissolved in 400 μl PBS.

Sun D., Evans L., Perrone F., Sokleva V., Lim K., Rezakhani S., Lutolf M., Zilbauer M, & Rawlins E.L. (2021). A functional genetic toolbox for human tissue-derived organoids. eLife, 10, e67886.

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Endoplasmic Reticulum Enrichment Protocol

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Cellular fractionation was performed using the ER enrichment kit (Novus Biologicals, Centennial, USA) according to product manual. In brief, 0.5 g cells were harvested and washed in PBS, then resuspended in 2 mL 1 X isosmotic homogenization buffer supplemented with protease inhibitor cocktail and homogenized thoroughly. The homogenate was centrifuged at 1000 g for 10 min at 4°C to get rid of nuclear, mitochondria, and cell debris. The supernatant was transferred to a new tube and centrifuged at 12,000 g for 15 min at 4°C. Then, the supernatant was transferred to a microcentrifuge tube and centrifuged at 90,000 g (Beckman Avanti J30I centrifuge with JS24 rotor) for 60 min at 4°C. The supernatant was kept as cytosol and the pellet (which contains the total ER fraction) was suspended in 1 X suspension buffer supplemented with protease inhibitor cocktail. Prepared cytosol and ER were kept at -80°C for further immunoblot.

Zhao J., He C., Fan X., Wang L., Zhao L., Liu H., Shen W., Jiang S., Pei K., Gao J., Qi Y., Liu Y., Zhao J., Zhang R., Lu C., Tong J, & Huai J. (2024). Tripeptidyl peptidase II coordinates the homeostasis of calcium and lipids in the central nervous system and its depletion causes presenile dementia in female mice through calcium/lipid dyshomeostasis-induced autophagic degradation of CYP19A1. Theranostics, 14(4), 1390-1429.

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