Ultrafree mc gv centrifugal filters

Manufactured by Merck Group

Sourced in United States

The Ultrafree-MC-GV centrifugal filters are a type of laboratory equipment designed for sample preparation and purification. They are used to separate and concentrate analytes from complex samples through a filtration process. The filters feature a hydrophilic PVDF membrane that allows for efficient sample recovery.

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Lab products found in correlation

Nsmol antibody ba kit, by Shimadzu (1 mentions) Aflibercept, by Santen (1 mentions) Gc rich pcr system, by Roche (1 mentions) 2 propanol, by Thermo Fisher Scientific (1 mentions) Lb media, by MP Biomedicals (1 mentions) Lc ms ultra grade formic acid fluka, by Merck Group (1 mentions) Sulfosalicylic acid, by Merck Group (1 mentions) Chelex 100 resin, by Bio-Rad (1 mentions) Hyclone, by Thermo Fisher Scientific (1 mentions)

6 protocols using ultrafree mc gv centrifugal filters

Isolation of OMVs from Campylobacter jejuni

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OMVs were isolated from C. jejuni strain 81176 using methods described by Elmi et al. (2012 (link)) and Chutkan et al. (2013 ).
Campylobacter jejuni 81176 strain was grown in the Mueller–Hinton broth under microaerobic conditions at 37 °C. An overnight culture was diluted 1:100 into 330 ml of fresh growth media and grown 16–18 h to a mid log phase. Briefly, the cells were pelleted twice, using centrifugation at 6000×g for 2 × 20 min at 4 °C. Supernatants were filtered through a 0.22-μm filter device to remove remaining cells. The filtrate was ultracentrifuged in Beckman L7-55 Ultracentrifuge at 150,000×g for 3 h at 4 °C, using a 50.2 Ti rotor. OMV preparations were plated on BA plates and incubated under microaerobic conditions to confirm the absence of viable bacteria.
The pellets were resuspended with Dulbecco’s phosphate-buffered saline (DPBS) or PBS and stored at −20 °C until needed. Before injecting into eggs, the OMV samples were sterilized by filtration using Ultrafree-MC-GV Centrifugal filters (Merck Millipore). A protein content of OMVs was measured by BCA assay.

Godlewska R., Kuczkowski M., Wyszyńska A., Klim J., Derlatka K., Woźniak-Biel A, & Jagusztyn-Krynicka E.K. (2016). Evaluation of a protective effect of in ovo delivered Campylobacter jejuni OMVs. Applied Microbiology and Biotechnology, 100(20), 8855-8864.

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Quantification of Aflibercept in Plasma

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An nSMOL Antibody BA kit was purchased from Shimadzu (Kyoto, Japan). Aflibercept was purchased from Santen Pharmaceutical (Osaka, Japan). Individual male and female control human plasma samples were obtained from Kohjin Bio (Saitama, Japan). P14R (14-Pro and Arg) internal standard synthetic peptides were purchased from Sigma-Aldrich (St. Louis, MO). Ultrafree-MC GV centrifugal filters (0.22 µm pore size) were procured from EMD Millipore (Billerica, MA). Other reagents, buffers, and solvents were purchased from Sigma-Aldrich and FUJIFILM Wako Pure Chemicals (Osaka, Japan).

Kimura N., Takahashi H., Sakamoto S., Yanagi Y., Maeshima N., Minamimoto A., Iwamoto N., Shimada T., Nagai R, & Aizawa K. (2022). Microvolume Analysis of Aflibercept in Aqueous Humor Using Mass Spectrometry. Translational Vision Science & Technology, 11(6), 7.

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Proteomic Profiling of Recombinant Proteins

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J.T. Baker Ultra LC/MS-grade acetonitrile (ACN) and water were purchased from VWR International (Radnor, PA). LC/MS Ultra-grade formic acid (Fluka) was purchased from Sigma-Aldrich (Saint Louis, MO, USA). Roswell Park.
Memorial Institute (RPMI) media (Gibco), pBAD Directional TOPO® Expression Kit, and ProBond™ Purification System were purchased from Invitrogcn (Carlsbad. CA, USA). DL- dithiothreitol (DTT), MgS0₄, and ¹³C₆, ¹⁵N₄-L-arginine hydrochloride were purchased from Sigma-Aldrich (St. Louis, MO, USA). LB media (MP Biomedicals), GC-Rich PCR system (Roche), 4-(2-hydroxyethyl)-l -pipcrazineethancsulfonic acid (HEPES) at pH 8.4, 2-propanol (Fisher Chemical), and borate buffer at pH 9.5 (R1CCA Chemical) were purchased from Fisher Scientific (Pittsburgh, PA, USA). The Quick Blunting kit and restriction enzymes (EcoRI, EcoRV, HindIII, SphI) were purchased from New England Biosciences (Ipswich, MA, USA). 4-Fluoro-7-nitro-2,l,3-bcnzoxadiazole (NBDF) was purchased from Dojindo Laboratories (Rockville, MD, USA). Amicon Ultra 3 kDa MW cutoff centrifugal filters and Ultrafree-MC GV centrifugal filters were purchased from Millipore (Milford, MA, USA). Polypropylene auto-sampler tubes with PTFE/silicone septa were purchased from Waters (Milford, MA, USA).

Dalluge J.J., McCurtain J.L., Gilbertsen A.J., Kalstabakken K.A, & Williams B.J. (2015). Determination of agmatine using isotope dilution UPLC-tandem mass spectrometry: application to the characterization of the arginine decarboxylase pathway in Pseudomonas aeruginosa. Analytical and bioanalytical chemistry, 407(18), 5513-5519.

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Protein-Bound Amino Acid Quantification

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20 μL of media was mixed with 80 μL of water and 100 μL of 10% Sulfosalicylic acid (Sigma-Aldrich, St. Louis, MO) followed by incubation for 1 hour at 4°C and then centrifugation at 13,000 rpm (15 min; 4°C). No visible pellet was seen. Supernatant was filtered using Ultrafree-MC-GV centrifugal filters (Millipore, Burlington, MA) and used for amino acid analysis employing Biochrom 30 amino acid analyzer. Results are in terms of μM and were normalized to total protein content.

Francescone R., Vendramini-Costa D.B., Franco-Barraza J., Wagner J., Muir A., Lau A.N., Gabitova L., Pazina T., Gupta S., Luong T., Rollins D., Malik R., Thapa R.J., Restifo D., Zhou Y., Cai K.Q., Hensley H.H., Tan Y., Kruger W.D., Devarajan K., Balachandran S., Klein-Szanto A.J., Wang H., El-Deiry W.S., Vander Heiden M.G., Peri S., Campbell K.S., Astsaturov I, & Cukierman E. (2020). Netrin G1 promotes pancreatic tumorigenesis through cancer associated fibroblast driven nutritional support and immunosuppression. Cancer discovery, 11(2), 446-479.

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Amino Acid Quantification in Cell Lysates

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14.55 μg of cell lysate protein in RIPA buffer was brought to a total volume of 50 μL in water and was mixed with 50 μL of 10% Sulfosalicylic acid (Sigma-Aldrich, St. Louis, MO) followed by incubation for 1 hour at 4°C and then centrifugation at 13,000 rpm (15 minutes; 4°C). Supernatant was filtered using Ultrafree-MC-GV centrifugal filters (Millipore, Burlington, MA) and used for amino acid analysis employing the Biochrom 30 amino acid analyzer (Biochrom US, Holliston, MA). Results were provided in nmol/mg protein units (normalized to total protein content).
These results are available in Supplemental Tables 5-7.

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Optimized Molecular Biology Protocols

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The chemical reagents used here are molecular biology grade or higher. DNA primers and oligonucleotides were purchased from Operon MWG. All aqueous solutions were prepared with deionized, distilled, nuclease free water (HyClone, Thermo Scientific). For the experiments in the absence of divalent cations, nuclease free water was treated with the Chelex 100 Resin (Biorad) chelating resin and recovered with 0.2 μm Ultrafree–MC–GV Centrifugal Filters (Milipore). All the experiments are reproducible and repeated at least 2 times unless otherwise stated.

Gulen B., Petrov A.S., Okafor C.D., Vander Wood D., O’Neill E.B., Hud N.V, & Williams L.D. (2016). Ribosomal small subunit domains radiate from a central core. Scientific Reports, 6, 20885.

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