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Amicon ultra 100 k filter unit

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The Amicon Ultra 100 K filter unit is a centrifugal filter device designed for the concentration and desalting of protein solutions. The filter unit features a 100 kDa molecular weight cutoff membrane that allows the passage of small molecules while retaining larger proteins and macromolecules.

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Lab products found in correlation

Rpmi medium, by Thermo Fisher Scientific (1 mentions) Hek293ft cells, by Thermo Fisher Scientific (1 mentions) Lipofectamine 2000 reagent, by Thermo Fisher Scientific (1 mentions) Aav 293 cells, by Agilent Technologies (1 mentions)

Close spelling variants of this product

Amicon Ultra (740 citations) Amicon filters (181 citations) Amicon Ultra filters (132 citations) Amicon filter units (21 citations) Filter units (3 citations) Amicon 100 K (2 citations) Amicon Ultra Centrifugal Filters-100 K Units (2 citations)

3 protocols using amicon ultra 100 k filter unit

1

Dilution Series of Virus-Infected Cell Media

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Medium of A549 infected with Ad6-hT-GM at MOI = 100 vp/cell and non-infected A549 cells were collected 48 hpi (hours post-infection) and clarified by centrifugation for 10 min at 2800× g. It was filtered then through the Amicon Ultra 100 K filter unit (Millipore Burlington, MA, USA) to remove the virus. Two-fold dilutions of the culture medium in the range of 1:4 to 1:1024 were prepared for the experiment in RPMI medium (Invitrogen Waltham, MA, USA) supplemented with 10% fetal calf serum (HyClone, Logan, UT, USA).
Osipov I.D., Vasikhovskaia V.A., Zabelina D.S., Kutseikin S.S., Grazhdantseva A.A., Kochneva G.V., Davydova J., Netesov S.V, & Romanenko M.V. (2023). Development of Oncolytic Vectors Based on Human Adenovirus Type 6 for Cancer Treatment. Viruses, 15(1), 182.
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2

Production and Purification of Lentivirus and AAV

Cited 2 times
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Lentivirus was produced by co-transfection of established lentiviral plasmids20 (link) and packaging vectors (RRE, Rev, VSVG) into HEK-293FT cells (Invitrogen). Transfection was performed using Lipofectamine 2000 reagent (Invitrogen) following the manufacturer's protocol. 48–72 h after transfection, supernatants were collected and virus was concentrated following our previously described protocol32 (link). AAV was also generated as previously described63 (link). Briefly, AAV plasmid and helper constructs were co-transfected into HEK-293 cells using CaCl2; supernatant and cells were collected 48–72 h later. Virus was precipitated using (NH4)2SO4, then the precipitate was re-suspended and purified on an iodixanol gradient by ultracentrifugation. Further concentration of virus was done using an Amicon Ultra-100 K filter unit (Millipore).
Seshadri S., Faust T., Ishizuka K., Delevich K., Chung Y., Kim S.H., Cowles M., Niwa M., Jaaro-Peled H., Tomoda T., Lai C., Anton E.S., Li B, & Sawa A. (2015). Interneuronal DISC1 regulates NRG1-ErbB4 signalling and excitatory–inhibitory synapse formation in the mature cortex. Nature Communications, 6, 10118.
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3

AAV Production Using 293 Cells

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AAV-293 cells (Agilent Technologies) were transfected with AAV vectors (pAAV-LStopL-GFP-shRNAmirs) and helper vectors (pAd helper vector and pAAV2/5 packaging vector) following a standard CaCl2 method with some modifications (61 ). Transfected cells were harvested 48 hours later and lysed by three rounds of freeze-thaw cycles. The lysates were further sonicated and virus-containing fractions were precipitated by adding ammonium sulfate. Then, the viruses were enriched by OptiPrep™ density gradient ultracentrifugation at 60,000 rpm for 1.5 h, followed by concentration of viruses using Amicon Ultra-100 K filter unit (Millipore). Virus titers were estimated using a qPCR-based method to detect ITR sequence as described previously (50 (link)). The following primers and probe were used: AAV2 ITR F: 5’-GGAACCCCTAGTGATGGAGTT-3’, AAV2 ITR R: 5’-CGGCCTCAGTGAGCGA-3’, AAV2 ITR Probe: 5’-CACTCCCTCTCTGCGCGCTCG-3.’
Kano S.I., Choi E.Y., Dohi E., Agarwal S., Chang D.J., Wilson A.M., Lo B.D., Rose I.V., Gonzalez S., Imai T, & Sawa A. (2019). Glutathione S-transferases promote proinflammatory astrocyte-microglia communication during brain inflammation. Science signaling, 12(569), eaar2124.
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