Pro250 homogenizer

Manufactured by PRO Scientific

Sourced in United States

The PRO250 homogenizer is a versatile laboratory instrument designed for the efficient homogenization and emulsification of a wide range of sample materials. It features a high-speed motor and a robust stainless steel housing, ensuring reliable performance and durability.

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16 protocols using pro250 homogenizer

Isolation and Reverse Transcription of Total RNA

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Total RNA was obtained from the tibiae shaft using a modified two-step purification protocol employing homogenization (PRO250 Homogenizer, 10 mm×105 mm generator, PRO Scientific IN, Oxford CT) in Trizol (Invitrogen, Carlsbad, CA) followed by purification over a QIAGEN RNeasy column (QIAGEN, Valencia, CA). RNA integrity was monitored by running RNA on 1 % Agarose gel in 120 V for 30 min. Intact RNA has both clear 18S and 28S ribosomal RNA bands with 28S rRNA approximately twice as intense as the 18S rRNA band. RNA was reverse-transcribed to single-stranded cDNA by using High-Capacity cDNA Archive Kit (ABI, California). Gene expressions were presented as fold changes from PL group [3 (link)].

Yao W., Dai W., Jiang L., Lay E.Y., Zhong Z., Ritchie R.O., Li X., Ke H, & Lane N.E. (2015). Sclerostin-antibody treatment of glucocorticoid-induced osteoporosis maintained bone mass and strength. Osteoporosis international : a journal established as result of cooperation between the European Foundation for Osteoporosis and the National Osteoporosis Foundation of the USA, 27(1), 283-294.

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Curcuminoid Stabilized Phospholipid Nanoparticles

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To prepare stabilizer for CSP, 2.5 g L-α-phosphatidylcholine (P7443, Sigma, USA) and 3.42 g sucrose esters (Gemfont Corporation, Taiwan) were sequentially incorporated into 400 mL water. The mixed stabilizer materials were stirred at 25°C, and 40 g curcuminoid powder with curcumin, demethoxycurcumin, and bisdemethoxycurcumin (Toong Yeuan, Taiwan) were added to form a 10 % curcuminoid aqueous solution. This non-homogenously mixed solution was subjected to a high-speed homogenization pretreatment from 4,000 to 6,000g for 10 minutes using a PRO250 homogenizer (PRO Scientific, USA). Next, a nano-grade wet grinder (Netzsch-Fein mahltechnik GmbH, Germany) carried on yttria-stabilized tetragonal zirconia for circulation milling with 0.2 mm beads for 180 minutes to obtain the aqueous dispersion. The average diameter of un-nanosized curcuminoid was 5140±178 nm, and the average diameter of CSP was 59±1 nm. Finally, the nanosized CSP composed of curcumin (83.56%), demethoxycurcumin (14.13%) and bisdemethoxycurcumin (2.31%) was obtained. Right before oral administration, CSP was diluted into the drinking water at concentration 0.75 mg/mL. The vesicle control in this study contain the same stabilizer and went through the same preparation process without adding curcuminoid.

Tai Y.H., Lin Y.Y., Wang K.C., Chang C.L., Chen R.Y., Wu C.C, & Cheng I.H. (2018). Curcuminoid submicron particle ameliorates cognitive deficits and decreases amyloid pathology in Alzheimer’s disease mouse model. Oncotarget, 9(12), 10681-10697.

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Efficient Extraction of Mouse Pancreas RNA

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Mouse pancreas tissue lysate was obtained by quickly removing a piece of the pancreas from mice killed by CO₂ asphyxiation and snap freezing in liquid nitrogen. The frozen tissue was then homogenized in either RLT+ buffer for RNA or RIPA buffer supplemented with ethylenediamine tetraacetic acid–free protease inhibitor and PhosSTOP phosphatase inhibitor (Roche, South San Francisco, CA) by using a Pro 250 Homogenizer (Pro Scientific Inc, Oxford, CT). Lysate was then cleared by centrifugation and stored at –80°C. RNA was processed by using an RNEasy Plus kit (Qiagen, Valencia, CA) following the manufacturer’s protocol.

Halbrook C.J., Wen H.J., Ruggeri J.M., Takeuchi K.K., Zhang Y., Pasca di Magliano M, & Crawford H.C. (2016). Mitogen-activated Protein Kinase Kinase Activity Maintains Acinar-to-Ductal Metaplasia and Is Required for Organ Regeneration in Pancreatitis. Cellular and Molecular Gastroenterology and Hepatology, 3(1), 99-118.

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Extracting Bioactive Compounds from Freeze-Dried Açaí

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Samples of 0.20 ± 0.01 g of ground freeze-dried açaí were extracted (in duplicate) with 5 mL of acidified 70% v/v High-Pressure Liquid Chromatography (HPLC) grade methanol in 0.5% v/v HPLC grade acetic acid (HAc, Fisher Scientific, Pittsburgh, PA, USA). The mixtures were homogenized using a Pro 250 homogenizer (Pro Scientific Inc., Oxford, CT, USA) for 10 min and centrifuged (Sorvall RC-6 plus, Asheville, NC, USA) for 10 min at 5000 rpm. The supernatant was transferred to a 25 mL volumetric flask. The resultant pellet of açaí was further extracted twice and the extracts were combined and diluted to a final volume of 25 mL. Each final solution was filtered using a 0.2 μm polytetrafluoroethylene (PTFE) syringe filter (Fisher Scientific, Pittsburgh, PA, USA) before analysis.

Matta F.V., Xiong J., Lila M.A., Ward N.I., Felipe-Sotelo M, & Esposito D. (2020). Chemical Composition and Bioactive Properties of Commercial and Non-Commercial Purple and White Açaí Berries. Foods, 9(10), 1481.

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Panx3 Protein Expression Analysis in Mouse Tissues

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Intact IVDs and AF tissues were harvested from 2-month-old (8 ± 2 weeks) WT and Panx3^-/- mice for protein analysis. Human embryonic kidney 293T cells overexpressing mouse Panx3 (described in [21 (link)]) served as control. Total protein was harvested following tissue homogenization (PRO250 homogenizer, PRO Scientific, Oxford, CT, USA) and sonication (Sonic Dismembrator 100, Fisher Scientific, Waltham, MA, USA) in Triton-based extraction buffer as previously described [21 (link)]. Following quantification using the bicinchoninic acid assay, 16 μg total protein were separated by gel electrophoresis on a 10% sodium dodecyl sulfate-polyacrylamide gel and transferred to a nitrocellulose membrane. Membranes were blocked for 1.5 h with 3% (w/v) bovine serum albumin in phosphate buffer saline (PBS) and incubated overnight at 4 °C with rabbit polyclonal anti-Panx3 primary antibody (1:1000; described in [21 (link)]). Membranes were washed and incubated for 45 min with IRDye 800CW goat anti-rabbit secondary antibody (1:10,000; LiCor, Lincoln, NE, USA; 925-32211) prior to visualization using Odyssey LiCor infrared imaging system (Lincoln, NE, USA). GAPDH was detected using a mouse monoclonal primary antibody (1:5000; Millipore Sigma, Burlington, MA, USA; MAB374), followed by incubation with IRDye 680RD goat anti-mouse secondary antibody (1:10,000; LiCor, Lincoln, NE, USA; 925-68070).

Serjeant M., Moon P.M., Quinonez D., Penuela S., Beier F, & Séguin C.A. (2021). The Role of Panx3 in Age-Associated and Injury-Induced Intervertebral Disc Degeneration. International Journal of Molecular Sciences, 22(3), 1080.

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DNA Extraction from Decayed Wood Samples

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Pooled dead wood samples of DC 1–4 were prepared by taking a spoonful of material from each sub-sample, adding sterile water (wood/water mixture of ~20 ml) and mixing using a Pro250 Homogenizer (Pro Scientific, Oxford, CT, USA). Extra water was removed by centrifuging samples at 9000 r.p.m. for 3 min. Samples were kept on ice before and after homogenisation, after which DNA extraction was started immediately. DNA extraction from 150 to 250 mg (fresh weight) samples was carried out using a PowerSoil DNA Isolation Kit (MoBio Laboratories, Carlsbad, CA, USA) according to the manufacturer’s instructions with the following modifications: cell lysis was performed using a MoBio vortex adapter for 15 min and incubating for 30 min at 60 °C.
DNA extraction from the soil samples and heavily decayed wood samples (decay class 5) were performed with the same commercial kit (PowerSoil DNA Isolation Kit, MoBio) following the standard protocol with slight modifications: cell lysis was performed using a Fast Prep homogenizer (MP Biomedicals, Santa Ana, CA, USA) three times for 20 s at 4 m s⁻¹ and incubating for 30 min at 65 °C.

Mäkipää R., Rajala T., Schigel D., Rinne K.T., Pennanen T., Abrego N, & Ovaskainen O. (2017). Interactions between soil- and dead wood-inhabiting fungal communities during the decay of Norway spruce logs. The ISME Journal, 11(9), 1964-1974.

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Pancreas Lysate Preparation Protocol

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For lysate preparation, the pancreas was removed from 12–14-week old mice and immediately homogenized in MLB buffer (25mM HEPES, pH 7.5, 150mM NaCl, 10mM MgCl₂, 1mM EDTA, 1% NP-40, and 10% glycerol) containing protease and phosphatase inhibitors (Roche) using a Pro 250 Homogenizer (Pro Scientific). For pancreas lysate immunoblots, 50 μg of protein was separated by denaturing SDS-PAGE, transferred to Immobilon P (Millipore) and visualized by chemiluminescence. Isolated acinar cells were washed once in PBS then lysed in MLB buffer containing protease inhibitors. 75 μg of protein was separated by SDS-PAGE, transferred to Immobilon P (Millipore) and visualized by chemiluminescence. Cell lines were washed once in PBS then lysed in RIPA buffer (150mM NaCl, 10mM Tris pH 7.2, 1% sodium deoxycholate, 1% Triton X-100, and 0.1% SDS) containing protease and phosphatase inhibitors. 25 μg of protein was separated by SDS-PAGE and visualized by chemiluminescence.

Wu C.Y., Carpenter E.S., Takeuchi K.K., Halbrook C.J., Peverley L.V., Bien H., Hall J.C., DelGiorno K.E., Pal D., Song Y., Shi C., Lin R.Z, & Crawford H.C. (2014). PI3K Regulation of RAC1 is Required for KRAS-induced Pancreatic Tumorigenesis in Mice. Gastroenterology, 147(6), 1405-1416.e7.

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Autophagy Gene Expression Analysis in Distal Tibiae

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Total RNA was obtained from the distal tibiae. Total RNA was isolated using a modified two-step purification protocol employing homogenization (PRO250 Homogenizer, 10mm × 105mm generator, PRO Scientific IN, Oxford CT) in Trizol (Invitrogen, Carlsbad, CA, USA). The autophagic focus RT-PCR gene pathway arrays and the primer sets were purchased from SABioscience, a Qiangen company, (Frederick, MD, USA). Each pathway gene array has pre-selected 96 genes that are related to autophagy pathways, housekeeping genes, and no primer or cDNA controls. Detailed gene information can be found at http://www.sabiosciences.com/RTPCR.php. We excluded genes with Ct values of > 35 for the pathway analysis [34 (link), 44 (link)].

Dai W., Jiang L., Lay Y.A., Chen H., Jin G., Zhang H., Kot A., Ritchie R.O., Lane N.E, & Yao W. (2015). Prevention of glucocorticoid induced bone changes with beta-ecdysone. Bone, 74, 48-57.

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Isolation of RNA from Mouse Tissues

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Organs from normal C57BL/6 mice were dissected and immediately frozen in liquid nitrogen and stored at −80 °C. To isolate RNA, 40–60 mg of frozen tissue specimens were cut in small pieces and transferred to a tube containing 1 mL of TRIzol Reagent (Invitrogen). Next, they were mechanically homogenized using a Pro250 homogenizer (PROScientific, Oxford, CT, USA) and total RNA was extracted according to the manufacturer’s instruction.

Polo-Generelo S., Torres B., Guerrero-Martínez J.A., Camafeita E., Vázquez J., Reyes J.C, & Pintor-Toro J.A. (2022). TGF-β-Upregulated Lnc-Nr6a1 Acts as a Reservoir of miR-181 and Mediates Assembly of a Glycolytic Complex. Non-Coding RNA, 8(5), 62.

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Wound Healing Gene Expression Analysis

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Total RNA was isolated from the fracture callus at day 14 using a modified two-step purification protocol employing homogenization (PRO250 Homogenizer, 10 mm × 105 mm generator, PRO Scientific IN, Oxford CT) in Trizol (Invitrogen, Carlsbad, CA), followed by purification over a Qiagen RNeasy column (Qiagen, Valencia, CA). RT-PCR gene pathway arrays focused on wound healing were purchased from SABiosciences (Frederick, MD) and included genes in the following categories: ECM Components (COL14A1, COL1A1, COL1A2, COL3A1, COL4A1, COL4A3, COL5A1, COL5A2, COL5A3, VTN), Remodeling Enzymes (Ctsg, Ctsk, Ctsl, F13a1, F3 (Tissue Factor), Fga (Fibrinogen), Mmp1a, Mmp2, Mmp7, Mmp9, Plat (tPA), Plau (uPA), Plaur (uPAR), Plg (plasminogen), Serpine1 (PAI-1 [Plasminogen activator inhibitor 1]), Timp1), Cellular Adhesion (Cdh1 (E-cadherin), Itga1, Itga2, Itga3, Itga4, Itga5, Itga6, Itgav, Itgb1, Itgb3, Itgb5, Itgb6), Cytoskeleton (Acta2 (a-SMA), Actc1, Rac1, Rhoa, Tagln), Inflammatory Cytokines & Chemokines (Ccl12, Ccl7 (Mcp-3), Cd40lg (Tnfsf5), Cxcl1, Cxcl11 (I-TAC/IP-9), Cxcl3, Cxcl5 (ENA-78/LIX), Ifng, Il10, Il1B, Il2, Il4, Il6) and Growth Factors (Angpt1, Csf2 (GM-CSF), Csf3 (GCSF), Ctgf, Egf, Fgf10, Fgf2, Fgf7, Hbegf (Dtr), Hgf, Igf1, Mif, Pdgfa, Tgfa, Tgfb1, Tnf, Vegfa). Data are presented as fold-differences from the placebo-treated (PBS) group for both sexes.

Yao W., Evan Lay Y.A., Kot A., Liu R., Zhang H., Chen H., Lam K, & Lane N.E. (2016). Improved Mobilization of Exogenous Mesenchymal Stem Cells to Bone for Fracture Healing and Sex Difference. Stem cells (Dayton, Ohio), 34(10), 2587-2600.

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