C57BL/6 mice were inoculated by the intranasal route with 10 µg of diABZI in 40 µl PBS with 5% DMSO. Vehicle-treated mice received 40 µl of 5% DMSO in PBS. 6 and 12 h post treatment, mice were sacrificed, and lungs were harvested. Lung tissues were snap-frozen in liquid nitrogen and transferred into 2 mL Lysing Matrix Tubes (MP Biomedicals) containing beads, followed by addition of 1 mL of TRIzol Reagent (Invitrogen). The samples were then homogenized using FastPrep-24™ Classic bead beating grinder and lysis system (MP Biomedicals). 900 μL of TRIzol Reagent was added to 100 μL of homogenized tissues for RNA extraction and subsequent RNAseq analysis.
Fastprep 24 classic bead beating grinder and lysis system
Manufactured by MP Biomedicals
Sourced in United States
The FastPrep-24™ Classic bead beating grinder and lysis system is a laboratory equipment designed for the rapid and efficient homogenization and lysis of a wide range of sample types. It utilizes high-speed bead beating to disrupt cells and tissues, enabling the release of cellular contents for downstream processing and analysis.
Automatically generated - may contain errors
Lab products found in correlation
Trizol reagent, by Thermo Fisher Scientific (1 mentions)
Lysing matrix tube, by MP Biomedicals (1 mentions)
Potato dextrose agar (pda), by Merck Group (1 mentions)
1.4 mm zirconium beads, by MP Biomedicals (1 mentions)
Halt protease and phosphatase inhibitor cocktail, by Thermo Fisher Scientific (1 mentions)
5 protocols using fastprep 24 classic bead beating grinder and lysis system
1
Intranasal diABZI Treatment in Mice
2
Quantifying Amyloid-beta Levels in Yeast
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BY4743 [pYEX.Aβ] and BY4743 [pYEX.BX] were cultured overnight and freshened for 2–3 h with fresh minimal media at 30 °C with shaking at 200× g. Following the freshening of the yeast transformants in the liquid media supplemented with auxotrophic requirements essential for growth, BY47473 [pYEX.Aβ] were treated with the combination of 15 μM trans-chalcone and 8 μM baicalein for two hours at 30 °C with shaking at 200× g. The study was performed along with the untreated control pYEX.Aβ and pYEX.BX transformants. After two hours of incubation, the yeast cells were harvested by centrifugation at 3000× g for 3 min, and cells were lysed using 0.5 mm beads in a FastPrep-24™ Classic bead beating grinder and lysis system (MP Biomedicals LLC., Irvine, CA, USA) at high speed for 1 min for 6 cycles. To avoid the heating of the tubes, each cycle of homogenization was followed by 1-min rest period on ice. The lysate obtained was analysed for the level of Aβ42 using MALDI-TOF mass spectrometry (Bruker Pty Ltd., Preston, VIC, Australia).
3
Fungal Isolation from Leaf Samples
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The same sets of samples used for Section 4.2 were used here. For each replicate sample, one leaf was excised from each of the three twigs and pooled before inoculation. The pooled leaves were surface-sterilized with 70% ethanol and 10% bleach, respectively, followed by three rinses in a large volume of sterilized distilled water (SDW), then blotted dry with a sterilized paper towel to remove epiphytes and associated DNA residues. They were then cut into small pieces and placed in a presterilized 2 mL microtube containing 1 mL SDW, 0.3 mL 1.4 mm Zirconium beads (MP Biomedicals, Santa Ana, CA, USA), and one ¼″ (6.35 mm) ceramic sphere (OPS Diagnostics, Lebanon, NJ, USA). Leaf pieces in the tubes were homogenized for 3 × 30 s at speed 6 on FastPrep-24™ Classic bead beating grinder and lysis system (MP Biomedicals, Santa Ana, CA, USA). An 0.3-mL aliquot of the resultant suspension was plated on potato dextrose agar (PDA, Sigma-Aldrich, St. Louis, MO, USA) in each 9-cm replicate Petri dish. These dishes were incubated at 23 °C for 7–10 days.
4
Extraction and Fractionation of Amyloid-beta
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The cortex from the right hemisphere was homogenized at 10% (w/v) in TBS (50 mM Tris–HCl, 150 mM NaCl, pH 7.6) containing Halt™ Protease and Phosphatase Inhibitor Cocktail (Thermo Fisher Scientific) using the FastPrep-24™ Classic bead beating grinder and lysis system (MP Biomedicals). The homogenized cortical tissue was aliquoted in protein LoBind tubes and stored at − 80 °C until analysis.
For extraction of Aβ, the prepared homogenates were thawed on ice and centrifuged at 14,000 × g for 30 min at 4 °C. The supernatant was collected as the TBS-soluble fraction, aliquoted in protein LoBind tubes, and stored at − 80 °C until analysis. The remaining pellet was re-suspended at 10% (v/w) in ice-cold 70% FA containing Halt™ Protease and Phosphatase Inhibitor Cocktail, sonicated on ice for 6 × 10 s, and centrifuged at 14,000 × g for 1 h at 4 °C. The supernatant was collected as the FA-soluble fraction, neutralized 1:20 in 1 M Tris-base at room temperature, aliquoted in protein LoBind tubes, and stored at − 80 °C until analysis.
For extraction of Aβ, the prepared homogenates were thawed on ice and centrifuged at 14,000 × g for 30 min at 4 °C. The supernatant was collected as the TBS-soluble fraction, aliquoted in protein LoBind tubes, and stored at − 80 °C until analysis. The remaining pellet was re-suspended at 10% (v/w) in ice-cold 70% FA containing Halt™ Protease and Phosphatase Inhibitor Cocktail, sonicated on ice for 6 × 10 s, and centrifuged at 14,000 × g for 1 h at 4 °C. The supernatant was collected as the FA-soluble fraction, neutralized 1:20 in 1 M Tris-base at room temperature, aliquoted in protein LoBind tubes, and stored at − 80 °C until analysis.
5
Fecal Transplant in Murine HCC Model
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Six-weeks-old recipient C57BL/6 mice were orally administered of an antibiotic cocktail, consisting of ampicillin (0.5 g/L), vancomycin hydrochloride (0.25 g/L), neomycin trisulfate salt hydrate (0.5 g/L), and metronidazole (0.5 g/L) in 1% sugar water for four weeks. Two weeks before fecal transplantation, eight-weeks-old donor C57BL/6 mice were given intrahepatic PBS or Hepa1-6 injection to establish the murine HCC model. Two weeks later, fecal pellets were collected from the donor HCC tumor-bearing mice or control mice. Feces pellets were mashed with the FastPrep-24™ Classic bead beating grinder and lysis system (MP Biomedicals), and resuspended in 2 ml saline solution to get a concentration of 60 mg fecal material per 200 μl saline. Recipient mice were orally gavaged with 60 mg fecal material. The gavage was performed on day 1, 2, 3, 4, 7 for a total of five doses. Another one week later, recipient mice were killed.
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