0.45 m membrane

Manufactured by Merck Group

Sourced in United States, Germany

The 0.45 µm membrane is a laboratory filtration product manufactured by Merck Group. It is designed for the removal of particulates, bacteria, and other microorganisms from various liquid samples. The membrane has a pore size of 0.45 micrometers, which allows for the effective filtration of a wide range of materials.

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Close spelling variants of this product

0.45 µm membrane filter (69 citations) 0.45 µm PVDF membranes (64 citations) 0.45 µm nitrocellulose membrane (9 citations) 0.45 µm nylon membrane filter (7 citations) PVDF) membranes (0.45 µm; (6 citations) 0.45 µm nylon syringe membranes (5 citations) Polyvinylidene difluoride membranes (0.45 µm (4 citations) Polyvinylidene fluoride membranes (0.45 µm; (3 citations) Poly(vinylidene difluoride) membrane (0.45 µm; (3 citations) Millipore membrane (0.45 µm) (3 citations)

31 protocols using 0.45 m membrane

Extraction and HPLC Analysis of Pomegranate Peel

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P. granatum peels were dried at 55 °C for 5 days. Five grams of dried and powdered (32-mesh) peels were extracted by the dynamic maceration process using a magnetic stirrer and water 1:100 (w/v) at 100 °C for 2 h. The extracts were filtered and concentrated in an airflow oven at 55 °C for 3 days. The yield of the extract obtained was calculated and expressed as a percentage.
For high-performance liquid chromatography (HPLC) analysis, the crude extract of P. granatum (AEPG) was dissolved in methanol:water (50:50, v/v) (1 mg/mL), filtered through a 0.45-µm membrane (Millipore, Merck, Billerica, MA, USA), and an aliquot of 10 μL was injected into the chromatographic system.

do Nascimento M.F., Cardoso J.C., Santos T.S., Tavares L.A., Pashirova T.N., Severino P., Souto E.B, & de Albuquerque-Junior R.L. (2020). Development and Characterization of Biointeractive Gelatin Wound Dressing Based on Extract of Punica granatum Linn. Pharmaceutics, 12(12), 1204.

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Establishing Stable PML-Silenced Cell Lines

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Complementary oligonucleotides against PML (siPML#1 and siPML#2), referred to the sequences of PML si1 and PML si3 in the reference [22] (link), were synthesized, annealed and ligated into pSIREN-RetroQ vector according to Knockout RNAi Systems User Manual (Clontech Laboratories, Inc. Mountain View, CA). These siRNA and non-specific siRNA (NC) plasmids with pSIREN-RetroQ or pLVX vector were co-transfected with packaging plasmids including pCMV-Gag pol, VSV-G or pMD2G and PSPA2 into HEK293T cells to produce retrovirus or lentivirus. Forty-eight hours later, the viral supernatants were collected, filtered through 0.45 µm membrane (Millipore) and respectively added into 293T cells incubated with the medium containing 1 µg/ml polybrene (santa cruz, sc-134220). Stably expressed cells were selected by 1 µg/ml puromycin after viral infection for 48 hours.

He W., Hu C.X., Hou J.K., Fan L., Xu Y.W., Liu M.H., Yan S.Y., Chen G.Q, & Huang Y. (2014). Microtubule-Associated Protein 1 Light Chain 3 Interacts with and Contributes to Growth Inhibiting Effect of PML. PLoS ONE, 9(11), e113089.

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Permeation of Curcumin and Quercetin through Porcine Nasal Mucosa

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The permeation tests were performed using vertical Franz-type diffusion cells (2.268 cm² surface area) equipped with freshly excised porcine nasal mucosa. The porcine nasal mucosa was obtained from a slaughterhouse authorized by the Ministry of Agriculture (Frigorífico Bonsul, Pelotas, RS, Brazil). Before the experiment, the porcine nasal mucosa was maintained for 30 min in SNF pH 6.4 [43 (link)].
The experiments were performed with the different developed nanocarriers. For the receptor solution, a SNF:PEG 400 (70:30, v/v; pH 6.4) mixture was used in order to maintain sink conditions at 37 ± 1 °C in a thermostatic bath with continuous magnetic stirring at 650 rpm for 12 h. Samples were withdrawn from the receptor compartment after 0.5, 2, 4, 8, 10, and 12 h of assay and immediately quantified by HPLC.
After 12 h, the porcine nasal mucosa was removed from the Franz apparatus, gently dried with a cloth, and washed with methanol to remove the excess formulation. To evaluate the retention of CUR and QU in the tissue, the exposed region of the porcine nasal mucosa was minced with a scalpel and placed in a volumetric flask with methanol, sonicated in an ultrasonic for 30 min, and stirred overnight. It was then filtered through a 0.45 µm membrane (Millipore Corporation, Billerica, MA, USA) and immediately quantified using HPLC.

Vaz G.R., Carrasco M.C., Batista M.M., Barros P.A., Oliveira M.D., Muccillo-Baisch A.L., Yurgel V.C., Buttini F., Soares F.A., Cordeiro L.M., Fachel F., Teixeira H.F., Bidone J., de Oliveira P.D., Sonvico F, & Dora C.L. (2022). Curcumin and Quercetin-Loaded Lipid Nanocarriers: Development of Omega-3 Mucoadhesive Nanoemulsions for Intranasal Administration. Nanomaterials, 12(7), 1073.

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Large-Scale Production of Soluble HLA-F Molecules

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Utilizing soluble HLA technology, CeLLine bioreactors (Integra Biosciences, Biebertal, Germany) were used for large scale production of recombinant sHLA-F*01:0x molecules [45 (link)]. Cell culture supernatant containing sHLA-F*01:0x molecules were harvested weekly, centrifuged and filtered through a 0.45 µM membrane (Millipore, Schwalbach, Germany) to remove cells and debris. Trimeric sHLA-F*01:0x molecules were purified using an N-hydroxy-succinimide (NHS)-activated HiTrap column (Life Technologies) coupled to the mAb W6/32. Purified proteins were verified quantitatively via an HLA class I-specific ELISA and qualitatively via SDS gel electrophoresis (Figure S1) and Western blot analysis Figure S2).

Hò G.G., Heinen F.J., Blasczyk R., Pich A, & Bade-Doeding C. (2019). HLA-F Allele-Specific Peptide Restriction Represents an Exceptional Proteomic Footprint. International Journal of Molecular Sciences, 20(22), 5572.

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Sulfentrazone Sorption Analysis in Soil

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The sorption analysis was performed using working solutions from a stock solution of 1000 mg L⁻¹ sulfentrazone (analytical standard, 92.01% purity) with concentrations of 1.0, 2.0, 5.0, 10.0, and 20.0 mg L⁻¹ in 0.01 mol L⁻¹ CaCl₂. The solutions for the Roundup Ready^®, Roundup Ultra^®, and Zapp Qi^® formulations were prepared in a 10 mg L⁻¹ N-(phosphonomethyl)glycine in 0.01 mol L⁻¹ CaCl₂. Into 50 mL capacity tubes were added 2.00 g soil, 5 mL sulfentrazone solution (varying with each concentration) and 5 mL of each formulation, according to each treatment proposed. In the treatment with sulfentrazone alone, 5 mL sulfentrazone solution (varying according to each concentration) and 5 mL 0.01 mol L⁻¹ CaCl₂ were added.
After this procedure, the tubes were sealed and agitated vertically at a controlled temperature (25 ± 2 °C) and 40 rpm for the equilibrium time previously determined. After stirring, the samples were centrifuged at 2260× g (3500 rpm) for seven minutes. A portion of the supernatant was filtered through a 0.45 µm membrane (Millipore^®) and transferred to 1.5 mL vials for the HPLC analysis.

Langaro A.C., Souza M.D., Pereira G.A., Barros J.P., da Silva A.A., Silva D.V., Passos A.B, & Mendonça V. (2020). Influence of Glyphosate Formulations on the Behavior of Sulfentrazone in Soil in Mixed Applications. Toxics, 8(4), 123.

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Bovine Rotavirus Isolation and Titration

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MA-104 cells (Rhesus monkey fetal kidney cells: ATCC, CRL-2378) were maintained by using Eagle’s Minimal Essential Medium (EMEM) (Nissui, Tokyo, Japan) supplemented with 10% fetal bovine serum (FBS) in our laboratory.
Representative bovine RVAs were isolated from each fecal sample according to the methods as previously described, with modifications [32 (link)]. Briefly, fecal samples were homogenized with serum-free EMEM and centrifuged at 2100× g for 15 min at 4 °C to remove debris. The supernatant was filtered through a 0.45 µm membrane (Millipore, Darmstadt, Germany) and treated with 10 µg/mL trypsin from bovine pancreas (Sigma Chemicals, MO, USA) at 37 °C for 1 h. Then, 200 µL of the treated supernatant were inoculated into monolayers of the MA-104 cells (2.2 × 10⁵ cells/mL) in glass tube (4 tubes per each isolate) and kept at 37 °C for 90 min. Thereafter, inoculum was removed from the cells, fed with EMEM containing 1.5 µg/mL trypsin, and incubated at 37 °C for 3 days. When cytopathic effects (CPEs) were observed by microscopy, the supernatant was harvested and repeatedly inoculated into newly prepared MA-104 cells until forth passage. The virus titers (a 50% tissue culture infective dose (TCID₅₀)/mL) were determined according to the method reported by Reed and Muench with fourth replicates [33 (link)].

Odagiri K., Yoshizawa N., Sakihara H., Umeda K., Rahman S., Nguyen S.V, & Suzuki T. (2020). Development of Genotype-Specific Anti-Bovine Rotavirus A Immunoglobulin Yolk Based on a Current Molecular Epidemiological Analysis of Bovine Rotaviruses A Collected in Japan during 2017–2020. Viruses, 12(12), 1386.

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Extraction of Phytochemicals from P. avium

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Hydroethanolic extracts and aqueous infusions of P. avium by-products were prepared according to the method previously described [13 (link)]. Firstly, leaves, stems, and flowers were reduced in a dried powder. For the hydroethanolic extracts, 1 g of the dried powder of each sample was dissolved in ethanol/water (50:50, v/v), and posteriorly sonicated for 30 min. After that, the hydroethanolic solutions were shaken at room temperature for 2 h and then again sonicated for 30 min. Finally, the extracts were filtered using a 0.45 µm membrane (Millipore, Bedford, MA, USA) in a vacuum system, evaporated under reduced pressure, freeze-dried, and stored at −20 °C until further use.
Regarding to aqueous infusions, 1 g of dried powder of each sample was subjected to an infusion (100 mL of water) at 100 °C for 3 min, according to the manufacturer’s instructions for daily herbal infusions. Then, infusions were filtered as described above to hydroethanolic extracts, freeze-dried, and stored at −20 °C until further analysis. The extraction yields were reported in a previous work [13 (link)].

Nunes A.R., Flores-Félix J.D., Gonçalves A.C., Falcão A., Alves G, & Silva L.R. (2022). Anti-Inflammatory and Antimicrobial Activities of Portuguese Prunus avium L. (Sweet Cherry) By-Products Extracts. Nutrients, 14(21), 4576.

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Extraction and Quantification of Plant Flavonoids

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Fresh plant materials, after being cleaned and dried under shady conditions, were dried at 75 °C for 48 h, and then powdered and filtered through a 40-mesh screen. The dried samples (1.00 g) were separately extracted with 60% ethanol (25 mL) for 2 h at 50 °C. Then, ultrasound-assisted extraction was performed for 20 min, the extraction processes were repeated twice. Finally, the mixture was filtered via a vacuum suction filter pump, and the extract solutions, which would be used to measure the total flavonoids content, were collected. Eight mL of extract solution was extracted twice with petroleum ether for removing the chlorophyll, and the residue solution was concentrated to dryness by evaporation on a rotary evaporator, and then dissolved with ethanol. Before testing the solutions were filtered through a 0.45 µm membrane (Millipore, Billerica, MA, USA). Samples for HPLC analysis were then prepared.

Wang X., Cao J., Wu Y., Wang Q, & Xiao J. (2016). Flavonoids, Antioxidant Potential, and Acetylcholinesterase Inhibition Activity of the Extracts from the Gametophyte and Archegoniophore of Marchantia polymorpha L.. Molecules, 21(3), 360.

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Bacterial Viability Measurement Protocol

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At the start and completion of each assay, E. coli concentration was measured by the Membrane Filtration (MF) technique using a 0.45 µm membrane (Millipore SAS, Billerica, MA, USA) and Brilliance (Oxoid Ltd., Basingstoke, UK) or mEndo (Becton, Dickinson and company) media followed by incubation at 37 °C for 24 h. This step was performed to obtain CFUs before and after the assay and to ensure bacterial viability and culturability.

Hesari N., Kıratlı Yılmazçoban N., Elzein M., Alum A, & Abbaszadegan M. (2017). A Strategy to Establish a Quality Assurance/Quality Control Plan for the Application of Biosensors for the Detection of E. coli in Water. Biosensors, 7(1), 3.

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Phospholipase C-mediated release of MASP49 and trans-sialidase

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CD-tryps (100 × 10⁶) from the Ninoa strain were washed three times with PBS and resuspended in 500 µL of DMEM without FBS. The parasites were incubated for 2 h at 37 °C with or without an addition of 4 U of PLC from Bacillus cereus (Sigma, USA) [33 (link)]. The supernatants were collected and centrifuged at 12,800× g for 15 min and filtered through a 0.45 µm membrane (Millipore, Burlington, MA, USA). Proteins from supernatants were quantified using Bio-Rad’s DC protein assay (Bio-Rad, Hercules, CA, USA), and 10 mg were loaded into 12% SDS-PAGE. Western blotting was performed as described above using anti-MASP49 serum (diluted 1:500) or anti-trans-sialidase antibodies (α-TS) (diluted 1:100): a gift from Dr. Sergio Schekman, Scola Paulista do Medicina, Brazil.

Espinoza B., Martínez I., Martínez-Velasco M.L., Rodríguez-Sosa M., González-Canto A., Vázquez-Mendoza A, & Terrazas L.I. (2023). Role of a 49 kDa Trypanosoma cruzi Mucin-Associated Surface Protein (MASP49) during the Infection Process and Identification of a Mammalian Cell Surface Receptor. Pathogens, 12(1), 105.

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