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Amicon ultra 0.5 ml centrifugal

Manufactured by Merck Group
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The Amicon® Ultra-0.5 mL centrifugal device is a laboratory filtration tool designed for the concentration and purification of macromolecules. It features a regenerated cellulose membrane with a specified molecular weight cut-off to facilitate the separation of desired molecules from complex solutions through centrifugation.

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Lab products found in correlation

Ultracel 30 membrane, by Merck Group (1 mentions) Ultracel 10 membrane, by Merck Group (1 mentions) His spintrap kit, by GE Healthcare (1 mentions) Masslynx 4, by Waters Corporation (1 mentions) Qubit protein assay, by Thermo Fisher Scientific (1 mentions) Cyclohexane, by Merck Group (1 mentions) S1805 photoresist, by MicroChem (1 mentions) Fluorescein isothiocyanate (fitc), by Thermo Fisher Scientific (1 mentions) Dimethyl sulfoxide (dmso), by Merck Group (1 mentions) Sodium hydroxide (naoh), by Mallinckrodt (1 mentions) Sodium bicarbonate, by Merck Group (1 mentions) Hepes, by Merck Group (1 mentions)

Close spelling variants of this product

Amicon® Ultra-0.5 ml centrifugal filter unit Amicon Ultra 0.5-mL Centrifugal Filters Ultracel 3K Amicon Ultra −0.5 mL Centrifugal Filter Units 100K Amicon Ultra-0.5 mL 10K centrifugal filters Amicon Ultra-0.5 ml Centrifugal Filters, 100,000 MWCO Amicon 30K Ultra-0.5 mL Centrifugal Filters Amicon 0.5 mL cellulose ultra centrifugal filters Amicon Ultra-0.5 mL centrifugal filter device Amicon Ultra-0.5 ml 3 K Centrifugal Filter Amicon Ultra-0.5 mL centrifugal filters

6 protocols using amicon ultra 0.5 ml centrifugal

1

Protein Binding Assay for SK14-061a

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Protein binding assays were performed following a previous reported procedure, with minor modifications [8 (link)]. In short, pooled human plasma, 600 μM HSA and 45 μM AGP were spiked with an aqueous solution of SK14-061a to produce a final SK14-061a concentration of 500 ng/mL, respectively. The unbound drug fraction was determined by ultrafiltration. Ultrafiltration was carried out using an Amicon® Ultra-0.5 mL centrifugal filter unit with an Ultracel®-30 membrane (Merck Millipore Company, MA). Samples of 500 μL were centrifuged at 14,000 g for 10 min. The SK14-061a concentration were determined by LC-MS without the SPE method.
Hashimoto M., Taguchi K., Ishiguro T., Kohgo S., Imoto S., Yamasaki K., Mitsuya H, & Otagiri M. (2018). Pharmacokinetic properties of a novel inosine analog, 4′-cyano-2′-deoxyinosine, after oral administration in rats. PLoS ONE, 13(6), e0198636.
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2

Protein Purification via Ultrafiltration

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Culture supernatant was upconcentrated using Amicon® Ultra 0.5 mL centrifugal filter unit with ultracel-10 membrane (Merck Millipore). Purification of His-tagged mRFP was performed with a His SpinTrap kit (GE Healthcare).
Schalén M., Anyaogu D.C., Hoof J.B, & Workman M. (2016). Effect of secretory pathway gene overexpression on secretion of a fluorescent reporter protein in Aspergillus nidulans. Fungal Biology and Biotechnology, 3, 3.
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3

Native Mass Spectrometry Analysis of Proteins

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Purified proteins (and complexes) were buffer exchanged into 200 mM ammonium acetate solution at pH 8.5 using 10 kDa cut-off Amicon Ultra 0.5 ml centrifugal spin filters (Merck Millipore). Samples were centrifuged six times at 12,000 rpm for 5 min at room temperature. Protein concentrations were determined using a Qubit protein assay (ThermoFisher Scientific), and samples were diluted to a concentration of 3.2 μM for native mass spectrometry analysis. Protein samples were introduced into a first-generation Synapt QToF mass spectrometer (Waters Corp) using a nanoelectrospray gold-coated borosilicate glass capillary (prepared in house). Instrument parameters were as follows: capillary voltage 1.2 kV, sampling cone voltage 55 V, source offset 2 V, backing pressure 3.2 mbar, or 5.9 mbar, trap collision energy 8 eV, transfer collision energy 6 eV, and bias 12 V. CID was performed using a trap collision energy of 28 eV and a transfer collision energy of 26 eV. Native mass spectrometry data were processed using MassLynx 4.2 (Waters Corp), UniDec and in house software Amphitrite (38 (link), 39 ).
Bagnéris C., Senthil Kumar S.L., Baratchian M., Britt H.M., Assafa T.E., Thalassinos K., Collins M.K, & Barrett T.E. (2022). Mechanistic insights into the activation of the IKK kinase complex by the Kaposi’s sarcoma herpes virus oncoprotein vFLIP. The Journal of Biological Chemistry, 298(6), 102012.
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4

Recombinant 14-3-3σ Preparation for Native MS

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Purified recombinant 14-3-3σ was exchanged into 100 mM ammonium acetate (pH 6.8) using an Amicon Ultra 0.5 mL centrifugal concentrator (Merck Millipore) with successive dilutions and concentrations. The exchanged protein was stored at −20 °C until use. A working stock of 20 μM (monomer) was diluted immediately before native MS analysis. Lyophilised myoglobin (horse heart) was purchased from Sigma-Aldrich, diluted to a stock concentration of 60 μM in 100 mM ammonium acetate (pH 6.8) and stored at 4 °C prior to use.
Bellamy-Carter J., Mohata M., Falcicchio M., Basran J., Higuchi Y., Doveston R.G, & Leney A.C. (2021). Discovering protein–protein interaction stabilisers by native mass spectrometry. Chemical Science, 12(32), 10724-10731.
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5

Fabrication of Microfluidic Devices

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The following chemicals were ordered from Sigma (St. Louis, MO): CHES, hydroxypropyl cellulose (HPC; 106 Da), poly(ethylene glycol) diacrylate (PEGDA; 575 Da), inhibitor remover beads, DMSO, and benzoin methyl ether (BME). FITC was purchased from Life Technologies (Carlsbad, CA). PTB biomarkers were obtained from the following sources: Fer from EMD Millipore (Billerica, MA), LF from Sigma (St. Louis, MO), P1 and P2 from Biomatik (Wilmington, DE), CRF and FITC-labeled P1 from GenScript (Piscataway, NJ), and TNF from ProSpec (East Brunswick, NJ). All solutions were made with deionized water (18.3 MΩ cm) from a Barnstead EASYpure UV/UF system (Dubuque, IA). Methanol was purchased from Macron (Center Valley, PA), sodium hydroxide from Mallinckrodt Baker (Paris, KY), and sodium bicarbonate from Merck (Darmstadt, Germany). HEPES, cyclohexane, and Amicon ultra 0.5 mL centrifugal (3, 10, 30, and 50 kDa cutoff) filters were purchased from EMD Millipore. Zeonor 1060R cyclic olefin copolymer (COC) was purchased in 1 and 2 mm thick sheets (Zeon Chemicals; Louisville, KY) cut to 2.2 x 5 cm pieces with an industrial bandsaw. Silicon wafers were from Fairchild (Phoenix, AZ) and S1805 photoresist came from MicroChem (Westborough, MA).
Nielsen A.V., Nielsen J.B., Knob R., Sahore V, & Woolley A.T. (2018). Microchip Electrophoresis Separation of a Panel of Preterm Birth Biomarkers. Electrophoresis, 39(18), 2300-2307.
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6

Plasma Protein Binding Determination Protocol

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Plasma protein binding (PPB) studies were conducted using a modified ultra-filtration technique [10 (link)]. Briefly, stock solutions were diluted with acetonitrile and spiked in blank rat plasma at three different concentrations: 50, 100, and 500 ng/mL for oncrasin-266 and oncrasin-72; and 25, 50, and 250 ng/mL for NSC-741908. The plasma was incubated at 37 °C for 10 min before being transferred to Amicon Ultra-0.5 mL centrifugal filters of 30kDa (EMD Millipore Corporation, Billerica, MA) for ultrafiltration at 20,800 x g for 15 min at 4°C. Filtrate and nonfiltrate plasma concentrations were spiked with IS and analyzed by LC-MS/MS.
Drug protein binding was calculated as PB = [1 ( Cu/Cp)]. Where PB represents protein binding, Cu is the unbound drug concentration, and Cp is the protein-bound drug concentration. Experiments were conducted in triplicate.
White L., Wu S., Ma J., Fang B, & Liang D. (2016). Simultaneous Determination and Validation of Oncrasin-266 and its Metabolites by HPLC-MS/MS: Application to a Pharmacokinetic Study. Journal of chromatography. B, Analytical technologies in the biomedical and life sciences, 1033-1034, 106-111.
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