Cellulose filter

Manufactured by Merck Group

Sourced in United States

Cellulose filters are a type of filtration media used in laboratory settings. They are made from purified cellulose fibers and are designed to remove particulates, microorganisms, and other contaminants from liquids or gases. Cellulose filters are commonly used in various applications, such as sample preparation, water treatment, and air filtration.

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Lab products found in correlation

Propidium iodide, by Thermo Fisher Scientific (3 mentions) Exoquick tc precipitation method, by System Biosciences (2 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions) Alginate, by FMC Biopolymer (1 mentions) Cellrox green, by Thermo Fisher Scientific (1 mentions) 1 hydroxybenzotriazole hydrate, by Merck Group (1 mentions) Rhodamine b isothiocyanate rbitc, by Merck Group (1 mentions) L glutamine, by Merck Group (1 mentions) Valinomycin, by Merck Group (1 mentions) Ultima gold, by PerkinElmer (1 mentions) Keratin azure, by Merck Group (1 mentions) Synergymx 96 well microplate reader, by Agilent Technologies (1 mentions) Waters breeze 2, by Waters Corporation (1 mentions) Rios 5, by Merck Group (1 mentions) Icap 6000, by Thermo Fisher Scientific (1 mentions) Qiaamp dna mini kit, by Qiagen (1 mentions)

15 protocols using cellulose filter

Cultivation and Cell-Free Supernatant Production

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The CMRP 4489 strain was activated on Luria–Bertani agar (LBA) (Neogen Corporation, USA) and incubated at 28 °C for 24 h. For the preparation of the pre-inoculum, colonies were suspended in saline solution (0.85% sodium chloride, w/v), and the concentration was adjusted according to the 0.5 McFarland scale until reaching approximately 1.5 × 10⁸ CFU/mL. For the preparation of the inoculum, 30 µL of pre-inoculum were inoculated separately in 125 mL Erlenmeyer flasks containing 30 mL of two culture media (CM1 or CM2) and incubated at 28 °C for 24 h at 125 rpm (Orbital shaker—Thoth 6430B, Brazil). Afterward, a 1% aliquot of the final volume (v/v) was transferred to a 1000 mL Erlenmeyer flask containing 400 mL of CM1 or CM2:
CM1–g/L: tryptone 10.0; yeast extract 5.0; NaCl 5.0; pH 7.1.
CM2–g/L: glucose 20.0; tryptone 12.4; NaCl 5.0; K₂HPO₄·3H₂O 1.5; MnSO₄·H₂O, 0.04; FeSO₄·7H₂O, 1.67; MgCl₂ · 6H₂O, 1.22; pH 7.1.
All culture media were incubated at 28 °C for 72 h at 150 rpm (Orbital shaker—Thoth 6430B, Brazil). Next, the fermentations were centrifuged for 10 min at 8860 × g at 4 °C (Hitachi, CR21G Himac, Japan) to obtain the cell-free supernatants (CFS). All CFS were sterilized by filtration through a cellulose filter with a pore size of 0.22 μm (Millipore, USA), thus obtaining CFS-CM1 and CFS-CM2.

Baptista J.P., Teixeira G.M., de Jesus M.L., Bertê R., Higashi A., Mosela M., da Silva D.V., de Oliveira J.P., Sanches D.S., Brancher J.D., Balbi-Peña M.I., de Padua Pereira U, & de Oliveira A.G. (2022). Antifungal activity and genomic characterization of the biocontrol agent Bacillus velezensis CMRP 4489. Scientific Reports, 12, 17401.

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Isolation of Small Extracellular Vesicles

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After selection with 4 µg/mL blasticidin-S, HuDe fibroblasts were further incubated for 72 h in serum free medium containing 4 µg/mL blasticidin-S to avoid any contamination by FBS lipoproteins. Then, medium was collected and centrifuged to remove cells, cell debris and large EVs (300× g, 10 min; 2000× g, 10 min), adding a filtration step at 0.22 µm with cellulose filter (Millipore, Burlington, MA, USA) to enrich for small EVs. Vesicles were isolated by polymer co-precipitation using Exoquick-TC precipitation method (System Biosciences, Palo Alto, CA, USA) as previously described [12 (link)]. Pelleted EVs were stored at −80 °C in PBS. EV protein content was determined by the Bradford method, using bovine serum albumin as standard.

Sagini K., Urbanelli L., Costanzi E., Mitro N., Caruso D., Emiliani C, & Buratta S. (2018). Oncogenic H-Ras Expression Induces Fatty Acid Profile Changes in Human Fibroblasts and Extracellular Vesicles. International Journal of Molecular Sciences, 19(11), 3515.

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Alginate-Based Biomaterial Synthesis

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Alginate (MW ≈ 250000 g mol⁻¹) was obtained from FMC Biopolymer. Cellulose filter was purchased from Millipore Inc. 2-(N-morpholino)ethanesulfonic acid (MES), potassium permanganate (KMnO₄), bovine serum albumin (BSA), poly(vinyl alcohol) (PVA, MW = 9000−10000), epigallocatechin gallate (EGCG), fluorescein amine, 1-hydroxybenzotriazole hydrate, rhodamine B isothiocyanate (RBITC), and L-glutamine were purchased from Sigma-Aldrich. PLGA (lactic:glycolic = 50:50, MW = 6000−10000 g mol⁻¹) was purchased from LACTEL. H₂O₂ (30% solution) was purchased from Macron Fine Chemicals. 1-Ethyl-3-(dimethylaminopropyl) carbodiimide hydrochloride, Pierce BCA Protein Assay Kit, Pierce Quantitative Peroxide Assay Kit, CellROX Green, propidium iodide, and Penicillin-Streptomycin were purchased from Thermo Scientific.

Seo Y., Leong J., Teo J.Y., Mitchell J.W., Gillette M.U., Han B., Lee J, & Kong H. (2017). Active Antioxidizing Particles for On-Demand Pressure-Driven Molecular Release. ACS applied materials & interfaces, 9(41), 35642-35650.

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Extracellular Vesicle Isolation from Ras-Transformed Fibroblasts

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HuDe fibroblasts transfected with H-RasV12 or with pcDNA6 were selected as described above. Before EVs recovery, cells were incubated for 72 hrs in serum free medium containing 4 μg/ml blasticidin-S to avoid any contamination by FBS lipoproteins that could affect EVs lipidomic analysis. Cells were counted in a haemocytometer and their viability was estimated as above. Medium was collected and underwent serial centrifugation steps to remove cells, cell debris and large EVs (300 × g, 10 min; 2,000 × g, 10 min), plus a filtration step at 0.22 μm with cellulose filter (Millipore), to enrich for small extracellular vesicles. EVs were isolated by polymer co-precipitation using Exoquick-TC^™ precipitation method (System Biosciences), according to the manufacturer’s instructions. Pelleted EVs were resuspended in PBS and stored at −80°C. Protein content was determined by the Bradford method, using bovine serum albumin as standard.

Buratta S., Urbanelli L., Sagini K., Giovagnoli S., Caponi S., Fioretto D., Mitro N., Caruso D, & Emiliani C. (2017). Extracellular vesicles released by fibroblasts undergoing H-Ras induced senescence show changes in lipid profile. PLoS ONE, 12(11), e0188840.

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Proteoliposome Cystine Uptake Assay

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Proteoliposomes were pelleted and resuspended in INSIDE buffer (10 mM MES-NaOH, 120 mM K-acetate, 2 mM MgSO₄ at pH 6.2). Four rounds of freeze-thaw were followed by extrusion through a 0.4 μm polycarbonate membrane. To reduce the volume, vesicles were harvested again at 18 °C and resuspended in INSIDE buffer.
Starting each individual assay, vesicles containing 2 μg protein were diluted into 250 μl OUTSIDE buffer (10 mM MES-NaOH, 120 mM NaCl, 2 mM MgSO₄ at pH 6.2) also containing 48 μM l-cystine, 2 μM radiocarbon-labelled ¹⁴C-l-cystine (specific activity 0.2 Ci mmol⁻¹, Hartmann Analytic) as well as 10 μM valinomycin (Merck). Assays were performed at 20 °C. 45 μl fractions were removed at different timepoints, diluted into 2 ml of OUTSIDE buffer and vesicles were immediately isolated on a 0.22 μm cellulose filter (Merck) using a vacuum manifold. Filters were washed twice with 2 ml OUTSIDE buffer and subsequently transferred into Ultima Gold (PerkinElmer) scintillation liquid. Remaining radioactivity stemming from the inside of the vesicles was measured using a Wallac scintillation counter.

Löbel M., Salphati S.P., El Omari K., Wagner A., Tucker S.J., Parker J.L, & Newstead S. (2022). Structural basis for proton coupled cystine transport by cystinosin. Nature Communications, 13, 4845.

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Soil Moisture Determination and Nitrate Amendments

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Soil moisture was determined gravimetrically by drying 5-10g soil at 60ºC for minimum 24h (ASTM D2216-10, 2010). Prior to incubation, soils were supplemented with NO3 -and ammonium by flooding and draining with a 2mM NH4NO3 solution e.g. (Samad et al., 2016a; Liu et al., 2010; Qu et al., 2014) . NO3 -supplied N for denitrification while ammonium acted as a preferential assimilatory N source. For this, an 80g dry weight equivalent of soil was placed in a 500mL Sterafil Filter Holder (Merck, Burlington, MA, USA) and flooded with 300mL of 2mM NH4NO3 (sufficient volume to dilute endogenous NO3 -) . After 15min the solution was drained through a 0.2µM cellulose filter (Merck) with 1.2µM glass-fibre pre filter (Merck) using a vacuum manifold. Soils were mixed and a subsample was taken for overnight moisture content analysis (5g, as above). The remainder of the soils was stored overnight in funnels covered with aluminum foil before use in incubation experiments the next day.

Highton M.P., Bakken L.R., Dörsch P., Wakelin S., de Klein C.A.M., Molstad L, & Morales S.E. (2020). Soil N2O emission potential falls along a denitrification phenotype gradient linked to differences in microbiome, rainfall and carbon availability. Soil biology & biochemistry, 150.

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Keratinase Activity Assay Protocol

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Keratinase activity assay was based on the method of Jaouadi et al. [40 (link)], with slight modification. The reaction mixture contained 0.5 mL of 10 g/L of keratin azure (Sigma-Aldrich, St. Louis, MO, USA) in 0.1 M Tris-HCl buffer, pH 7.5, and 0.5 mL of suitably diluted crude enzyme solution. The mixture was incubated at 37 °C for 1 h, with shaking at 220 rpm; after that, the reaction was stopped by placing the assay mixture in ice-cooled water for 10 min. The unutilized substrates were removed by centrifugation at 15,000× g for 10 min, and subsequently filtered (Millipore cellulose filters; 0.45 μm). The azo dye released in the filtrate was determined at 595 nm, using a SYNERGYMx 96 well microplate reader (BioTek Instrument Inc., Winooski, VT, USA). The control was treated at the same condition which contained the enzyme solution and buffer without the substrate. One keratinase unit was defined as the amount of enzyme causing an increase in absorbance of 0.01 per hour under the standard assay condition.
The total protein concentration was estimated by using the Bradford method [41 (link)], with bovine serine albumin as a standard protein. The respective assays were done in triplicate, and the results presented were mean plus standard deviation.

Nnolim N.E., Okoh A.I, & Nwodo U.U. (2020). Bacillus sp. FPF-1 Produced Keratinase with High Potential for Chicken Feather Degradation. Molecules, 25(7), 1505.

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Bacterial Extracts for Cell Culture

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Clinical isolates of Escherichia coli (Gram⁻ HMB-PP⁺ vit.B2⁺), Klebsiella pneumoniae (Gram⁻ HMB-PP⁺ vit.B2⁺), Pseudomonas aeruginosa (Gram⁻ HMB-PP⁺ vit.B2⁺), Corynebacterium striatum (Gram⁺ HMB-PP⁺ vit.B2⁺), Listeria monocytogenes (Gram⁺ HMB-PP⁺ vit.B2⁻), Staphylococcus aureus (Gram⁺ HMB-PP⁻ vit.B2⁺), Streptococcus pneumoniae (Gram⁺ HMB-PP⁻ vit.B2⁻) and Enterococcus faecalis (Gram⁺ HMB-PP⁻ vit.B2⁻) were grown in LB broth, harvested at an OD₆₀₀ of 0.5-0.8, and sonicated in 1/10 (v/v) PBS, pH 8.0. Insoluble debris was removed by centrifugation, the supernatants were passed through 0.1 μm sterile filter units (Millipore), and the protein concentrations were determined using the BCA protein assay kit (Pierce). Low molecular weight fractions were obtained using cellulose filters with a molecular mass cut-off of 3 kDa (Millipore). Bacterial extracts were used in cell culture at dilutions corresponding to protein concentrations of the original samples (before 3 kDa filtration) of 60-100 µg/ml.

Liuzzi A.R., Kift-Morgan A., Lopez-Anton M., Friberg I.M., Zhang J., Brook A.C., Roberts G.W., Donovan K.L., Colmont C.S., Toleman M.A., Bowen T., Johnson D.W., Topley N., Moser B., Fraser D.J, & Eberl M. (2016). Unconventional Human T Cells Accumulate at the Site of Infection in Response to Microbial Ligands and Induce Local Tissue Remodeling. The Journal of Immunology Author Choice, 197(6), 2195-2207.

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Neurotransmitter Analysis in Mouse Brain

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Mouse brain tissues were collected and homogenized in 0.2 ml ice-cold perchloric acid (0.4 M), followed by centrifugation at 12,000× g for 20 min, 4°C. The supernatants were filtered through 0.22 mm Cellulose filters (Millipore, USA). The resulting solution was injected into the HPLC system for electrochemical detection (Model 5600A; Coularray Detector System, ESA, Chelmsford, MA, United States) [41 (link), 42 (link)]. The neurotransmitters analyzed included dopamine (DA), 3,4-dihydroxyphenylacetic acid (DOPAC), 5-hydroxytryptamine (5-HT), 5-hydroxyindoleacetic acid (5-HIAA), and homovanillic acid (HVA) and were expressed as μg/g protein. DA and 5-HT turnover rates were calculated as DOPAC/DA and 5-HIAA/5-HT ratios, respectively.

Gao Y., Ma L., Gao F., Sun Z., Zhang Z., Li Y, & Li R. (2021). Endogenous Estrogen Influences Predator Odor-Induced Impairment of Cognitive and Social Behaviors in Aromatase Gene Deficiency Mice. Behavioural Neurology, 2021, 5346507.

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Docetaxel Quantification and Encapsulation Efficiency

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Docetaxel was quantified using high-pressure liquid chromatography (HPLC) in a Waters Breeze 2 (Waters Technologies, Milford, MA, USA) and a C18 Gemini, 5 µM, 150 × 4.60 mm column, at 35 °C. The mobile phase was a mixture of methanol:water 70:30 (v/v). The flow rate, injection volume and wavelength detection were set at 1 mL.min⁻¹, 20 µL and 270 nm, respectively [79 (link)]. The encapsulation efficiency (% EE) of DTX by the nanoparticles was determined by the ultrafiltration–centrifugation method, using cellulose filters (10 kDa, Millipore). Briefly, the total amount (100%) of DTX in the NLC was determined (DTX_total) by diluting the samples in the mobile phase (n = 3). The amount of DTX in the filtrate (DTX_free) was quantified by HPLC and the percentage of encapsulated DTX was calculated according to equation 2. Drug loading, the amount of loaded DTX in relation to the total weight of the nanoparticles, was also calculated according to Equation (3) [10 (link)]:

% EE = \frac{DTXtotal - DTX free}{DTX total} \times 100

% D r u g L o a d i n g = \frac{w e i g h t o f e n c a p s u l a t e d D T X}{w e i g h t o f n a n o p a r t i c l e s} \times 100

de Carvalho F.V., Ribeiro L.N., de Moura L.D., Rodrigues da Silva G.H., Mitsutake H., Mendonça T.C., Geronimo G., Breitkreitz M.C, & de Paula E. (2022). Docetaxel Loaded in Copaiba Oil-Nanostructured Lipid Carriers as a Promising DDS for Breast Cancer Treatment. Molecules, 27(24), 8838.

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