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Freezone 2.5 lyophilizer

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The FreeZone 2.5 lyophilizer is a freeze drying system designed for laboratory use. It is capable of removing water from samples through the process of sublimation, which involves transforming frozen water directly into water vapor without passing through the liquid phase. The FreeZone 2.5 can accommodate sample volumes up to 2.5 liters.

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Lab products found in correlation

0.45 m multiscreen filter plate, by Merck Group (2 mentions) Wiley mill, by Thomas Scientific (1 mentions) Infinite 200 pro, by Tecan (1 mentions) Silver nitrate, by SD Fine Chemicals (1 mentions) Mini extruder, by Avanti Polar Lipids (1 mentions) Pi 4 5 p2, by Avanti Polar Lipids (1 mentions) Ethanol, by EKF Diagnostics (1 mentions) 1 2 dipalmitoyl sn glycero 3 phosphocoline dppc, by Avanti Polar Lipids (1 mentions)

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Lyophilizer (104 citations) FreeZone 2.5 (89 citations) FreeZone lyophilizer (8 citations) Freezone® 2.5 model 76530 lyophilizer (3 citations)

12 protocols using freezone 2.5 lyophilizer

1

Extraction and Preparation of Plant Extracts

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Following shipment and storage at −80°C, plant materials were dried at 35°C in a dehumidifying drier for 3 days and ground into fine powder (2 mm mesh size) with a Thomas Scientific Wiley Mill (Swedesboro, NJ). Retention vouchers of chopped and ground material were prepared for future reference. For the preparation of organic crude extracts, plant powder was macerated for 72 h in 1 L flasks at a 1:10 ratio (w/v) in 95% ethanol with daily agitation. Macerates were filtered, the marc collected, and residual plant material subjected to a repeat extraction in 95% ethanol. Solvent was removed by rotary evaporation at ≤40°C. Extracts were redissolved in deionized water (dH2O), shell frozen in a dry ice-acetone bath, and then lyophilized overnight on a Labconco FreeZone 2.5 Lyophilizer (Kansas City, MO). Dry extracts were stored in scintillation vials at −20°C.
Aqueous extracts were prepared by boiling 30–40 g of plant material (1:10 ratio w/v) for 20 min in dH2O on a hot plate, followed by centrifugation and vacuum filtration. Aqueous extracts were then evaporated on a rotary evaporator and subsequently lyophilized as described above. Organic and aqueous crude extracts of all the plant samples were dissolved in DMSO or dH2O, respectively, to a stock concentration of 10 mg mL−1 prior to testing in biological assays.
Khan M.F., Tang H., Lyles J.T., Pineau R., Mashwani Z.U, & Quave C.L. (2018). Antibacterial Properties of Medicinal Plants From Pakistan Against Multidrug-Resistant ESKAPE Pathogens. Frontiers in Pharmacology, 9, 815.
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2

Synthesis and Characterization of Ts-AgNPs

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Ts-AgNPs were synthesized and characterized as we have reported earlier [14 (link)]. Briefly, 1 mg/mL tryptone (pH 12) (SRL, Bangalore, India) was mixed with 1 mM of silver nitrate (SD fine chemicals, Mumbai, India) and the solution was kept for heating for 20 min in the dark till the colour changed to dark brown. The following day, the nanoparticles were collected by centrifugation (26,200×g) for 30 min and the pellets were dried using a Labconco FreeZone 2.5 lyophilizer. The formation of Ts-AgNPs was confirmed by observing the UV–Vis spectrum (Infinite® 200 PRO, Tecan, Switzerland). The core size and shape of the Ts-AgNPs were visualized using a transmission electron microscope. The hydrodynamic diameter and the stability of the synthesized Ts-AgNPs were checked using a Zetasizer Nano-ZS90 size analyser (Malvern Instruments Ltd, Worcestershire, UK). The Fourier-transform infrared (FTIR) spectral analysis was employed to detect the presence of different functional groups on the nanoparticles [14 (link)].
Pandey P., Meher K., Falcao B., Lopus M, & Sirisha V.L. (2022). Tryptone-stabilized silver nanoparticles’ potential to mitigate planktonic and biofilm growth forms of Serratia marcescens. Journal of Biological Inorganic Chemistry, 28(2), 139-152.
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3

Unilamellar Lipid Vesicle Preparation

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Lipids were stored at −20°C. POPC (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine, designated as PC) was purchased from Larodan Inc, DOPS (1,2-di-oleoyl-sn-glycero-3-phospho-l-serine, designated as PS) was obtained from NOF America Corporation, and PI(4,5)P2 (brain L-α-phosphatidylinositol-4,5-bisphosphate, designated as PIP2) was purchased from Avanti Polar Lipids. Lipids dissolved in chloroform were mixed in a glass tube. The lipid mixture was dried under a stream of nitrogen gas for 30 min, and residual solvent was removed under vacuum (Labconco Free Zone 2.5 lyophilizer) for 2 hours. The dried lipid film was then rehydrated in the HEPES buffer (50 mM HEPES, 150 mM NaCl, pH 7.4), followed by a brief sonication to homogenize the mixture. The resulting lipid suspension was then extruded 21 times through 200 nm pore-size polycarbonate membranes, using a Mini extruder (Avanti Polar lipids) to form unilamellar vesicles.
Zhang L., Wang Y., Dong Y., Pant A., Liu Y., Masserman L., Xu Y., McLaughlin RN J.r, & Bai J. (2022). The endophilin curvature-sensitive motif requires electrostatic guidance to recycle synaptic vesicles in vivo. Developmental cell, 57(6), 750-766.e5.
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4

Lipid-Oligonucleotide Hybrid Liposomes

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DPPC and cholesterol were purchased from Avanti Polar Lipids (Alabaster,
AL) while poloxamer L64 was purchased from Sigma-Aldrich Chemicals (St. Louis,
MO). DPPC, cholesterol and L64 were mixed at 5:3:7 weight ratios in the presence
of excess tertiary butanol (Thermo Fisher Scientific, Waltham, MA), frozen at
−80°C overnight and lyophilized with a FreeZone 2.5 lyophilizer
(Labconco, Kansas City, MO) and stored for up to 3 months at 20°C.
Fluorescent DPPC or DCL64 liposomes were prepared by adding lissamine rhodamine
B-labeled DPPC (Avanti Polar Lipids) to the lipid mixture at 5% weight ratio,
frozen and lyophilized.
An 18-base nontoxic oligonucleotide,
5’-AGATGAACTTCAGGGTCA-3’, conjugated with Cyanine3 (Cy3) at the
5’ end was purchased from Trilink Biotechnologies (San Diego, CA). Bases
1, 2, 17, 18 were made of 2’-O-methyl modified phosphodiesters to enhance
oligonucleotide stability while bases 3 to 16 were made of normal
phosphodiesters. Oligonucleotides were mixed with DCL64 at a 1:10 weight ratio
at room temperature. L64 and oligonucleotides were incubated for 30 min,
followed by incubation with DPPC for 10 min, then with cholesterol for 10 min.
The lipid oligonucleotide mixture was frozen overnight at −80°C
and lyophilized.
Ashizawa A.T., Holt J., Faust K., Liu W., Tiwari A., Zhang N, & Ashizawa T. (2019). Intravenously Administered Novel Liposomes, DCL64, Deliver Oligonucleotides to Cerebellar Purkinje Cells. Cerebellum (London, England), 18(1), 99-108.
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5

Preparation of Kalanchoe Plant Extracts

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A total of seven crude extracts were prepared, four from K. mortagei and three from K. fedtschenkoi (Table 4). Each extract represented a particular plant part or combination of parts, though extract creation was also guided by limitations in available plant biomass.
Dry, ground plant biomass was double macerated for 72 h each with either 80 or 95% ethanol at a 1:10 ratio (w/v). The extracts were agitated daily and then vacuum filtered. The aqueous extract (1509aq) was prepared as a decoction; the dry plant material was boiled with deionized water (dH2O) for 20 min and then filtered. After filtration the solvent was removed by rotary evaporation at ≤40°C. Extracts were redissolved in dH2O, shell frozen in a dry ice-acetone bath, and then lyophilized overnight on a Labconco FreeZone 2.5 Lyophilizer (Kansas City, MO, United States). Dry extracts were scraped into scintillation vials and stored at -20°C. Organic extracts were dissolved in DMSO and the aqueous extract was re-dissolved in dH2O to yield a stock concentration of 10 mg mL-1 for microbiological assays.
Richwagen N., Lyles J.T., Dale B.L, & Quave C.L. (2019). Antibacterial Activity of Kalanchoe mortagei and K. fedtschenkoi Against ESKAPE Pathogens. Frontiers in Pharmacology, 10, 67.
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6

Mulberry Leaves and Fruits: Botanical Identification

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Mulberry leaves and fruits (Morus alba breeding clones ‘Ukraińska,’ ‘Bistro,’ and ‘Żółwińska wielkolistna,’ and Morus bombycis ‘Kenmochi’) were collected in July 2018 in the area of Rzeszów (Rzeszów, Poland, 50°00’N 22°01’E). After botanical identification, done by experts from the Aeropolis Laboratory of Plant Biotechnology (University of Rzeszow, Rzeszów, Poland), the leaves were dried at room temperature, without exposure to sunlight. The fruits were lyophilized using a FreeZone 2.5 lyophilizer (Labconco, Kansas City, MO, USA). Dehydration was carried out for 48 h by heating the shelves to 30 °C at a normal pressure of 0.5 bar. Both leaves and fruits were ground by a laboratory grinder into the form of powder. Voucher specimens of the plant material were deposited in the department archive. For experiments, rape honey from the beekeeping 2018 season obtained from the ecological apiary localized in the Podkarpackie region (J. Bańkowski, Roźwienica, Poland), liquefied at 45 °C during 48 h in a laboratory dryer, was used.
Tomczyk M., Miłek M., Sidor E., Kapusta I., Litwińczuk W., Puchalski C, & Dżugan M. (2019). The Effect of Adding the Leaves and Fruits of Morus alba to Rape Honey on Its Antioxidant Properties, Polyphenolic Profile, and Amylase Activity. Molecules, 25(1), 84.
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7

Ultrasound-assisted Extraction of Plant Compounds

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The dry plant material was grounded in a laboratory mill (A11 IKA, Germany) to obtain a homogenous drug powder. Drug material (2 g) was extracted for 30 min with 20 mL of 50% (v/v) ethanol (Stanlab, Lublin, Poland) solution using an ultrasound-assisted method (U-504 Ultron, Moorpark, CA, USA). Subsequently, the extract was filtered through a paper filter. ethanol was removed under vacuum using a rotary evaporator (Heidolph G3, Heidolph, Schwabach, Germany) at 40 °C. The extracts were prepared in triplicates. Obtained ethanol-free water extract was frozen at −80 °C and lyophilised for 24 h using FreeZone 2.5 lyophilizer (Labconco, Kansas City, MO, USA). Lyophilised extracts were diluted in sterile water, shortly before the experiment, to the final concentrations of 125, 250, 500 and 1000 µg/mL.
Sowa P., Marcinčáková D., Miłek M., Sidor E., Legáth J, & Dżugan M. (2020). Analysis of Cytotoxicity of Selected Asteraceae Plant Extracts in Real Time, Their Antioxidant Properties and Polyphenolic Profile. Molecules, 25(23), 5517.
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8

Ethanol Extraction of Plant Biomass

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Dry, ground plant biomass was double macerated in 95% ethanol at a 1:10 ratio (w/v) for 72 h of maceration with daily agitation. The liquid was then decanted and vacuum filtered, then concentrated by rotary evaporation at ≤40°C. Extracts were redissolved in dH2O, shell frozen in a dry ice–acetone bath, and lyophilized overnight on a Labconco FreeZone 2.5 Lyophilizer (Kansas City, MO). Dry crude extract (Extract ID: 649) was scraped into scintillation vials and stored at −20°C until microbiological testing, at which time they were dissolved in DMSO to a stock concentration of 10 mg ml−1.
Pineau R.M., Hanson S.E., Lyles J.T, & Quave C.L. (2019). Growth Inhibitory Activity of Callicarpa americana Leaf Extracts Against Cutibacterium acnes. Frontiers in Pharmacology, 10, 1206.
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9

Triphala Extract Preparation for Research

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Triphala capsules were purchased from the Himalaya Drug Company, India. 20% (w/v) Triphala powder was suspended in Milli-Q water. This partially soluble sample was then heated to 65 °C for 30 min. The sample was then centrifuged at 13 000 rpm and the soluble fraction was lyophilized with the help of a Labconco Free Zone 2.5 lyophilizer, USA. Required quantities of the lyophilized powder were weighed and dissolved in phosphate buffer saline (PBS) (10 mM Na2HPO4, 1.8 mM KH2PO4, 137 mM NaCl, 2.7 mM KCl and 0.01% (w/v) sodium azide) pH 7.4. The pH of the Triphala solution was adjusted to 7.4 before utilizing the sample for experiments.
Bopardikar M., Bhattacharya A., Rao Kakita V.M., Rachineni K., Borde L.C., Choudhary S., Koti Ainavarapu S.R, & Hosur R.V. (2019). Triphala inhibits alpha-synuclein fibrillization and their interaction study by NMR provides insights into the self-association of the protein. RSC Advances, 9(49), 28470-28477.
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10

Fabrication of DCL64 Liposomal Nanocarriers

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1,2-Dipalmitoyl-sn-glycero-3-phosphocoline (DPPC) and cholesterol were purchased from Avanti Polar Lipids and poloxamer L64 was purchased from Sigma-Aldrich. DCL64 liposomes were prepared by mixing DPPC, cholesterol, and poly(ethylene glycose)-block-poly(propylene glycol)-block-poly(ethylene glycol) L64 (poloxamer L64) at a 5:3:7 weight ratio at room temperature. DNAzyme was mixed with DCL64 at a 1:10 weight ratio at room temperature. All DCL64 liposomes were prepared in Wheaton glass vials and mixed with excess tert-butanol at room temperature prior to freezing at − 80 °C overnight. The DCL64 liposomes were then lyophilized overnight, using a FreeZone 2.5 lyophilizer (Labconco), until a thin film had formed, and there was no evidence of liquid within the glass vial. DCL64 liposomes not needed for use immediately were stored at − 20 °C. The DCL64 liposomes were reconstituted with sterile 1 × PBS to the desired concentration immediately before use.
Zhang N., Bewick B., Schultz J., Tiwari A., Krencik R., Zhang A., Adachi K., Xia G., Yun K., Sarkar P, & Ashizawa T. (2021). DNAzyme Cleavage of CAG Repeat RNA in Polyglutamine Diseases. Neurotherapeutics, 18(3), 1710-1728.
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