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Kv500 filter

Manufactured by Schott
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The KV500 filter is a laboratory equipment product that serves as a filtration device. It is designed to remove particulates and impurities from liquids or gases passing through it.

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Sx 20mv stopped flow apparatus, by Applied Photophysics (3 mentions)

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KV500 cut-off filter (3 citations)

3 protocols using kv500 filter

1

Rapid Kinetic Binding of RF1-GAQ

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Rapid kinetic experiments were performed on an SX-20MV stopped-flow apparatus (Applied Photophysics, Leatherhead, UK), by rapidly mixing equal volumes (60 µl) of reactants at 37 °C in buffer A. Binding of RF1-GAQ was monitored by mixing termination complex labelled with fluorescein (50 nM) with RF1-GAQQSY9 (300 nM). Fluorescein was excited at 470 nm and monitored after passing a KV500 filter (Schott, Mainz, Germany).
Graf M., Huter P., Maracci C., Peterek M., Rodnina M.V, & Wilson D.N. (2018). Visualization of translation termination intermediates trapped by the Apidaecin 137 peptide during RF3-mediated recycling of RF1. Nature Communications, 9, 3053.
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2

Rapid Kinetics of Ribosome Release

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Rapid kinetics measurements were performed on an SX-20MV stopped-flow apparatus (Applied Photophysics, Leatherhead, UK). Experiments were performed by rapidly mixing equal volumes (60 μl) of f[3H]Met-tRNAfMet(Flu)-carrying PreHC (0.05 μM), preincubated with Api137 for 2 min at room temperature and RF1Qsy (0.15 μM) at 37°C. Fluorescein was excited at 470 nm and fluorescence emission was monitored after passing a KV500 filter (Schott). Time courses were evaluated by fitting using exponential functions by GraphPad Prism software. Dissociation rates (koff) were determined by chase experiments: PreHCflu (0.05 μM) was preincubated with 0.15 μM RF1Qsy to generate PostHCflu in the absence or presence of 1 μM Api137. PreHC was then rapidly mixed with a 10-fold excess of unlabeled RF1 and RF3·GTP (1 mM); pyruvate kinase (0.1 mg/ml), and phosphoenol pyruvate (3 mM) were present in both syringes. The increase of fluorescence upon dissociation of RF1Qsy was monitored as described above.
Florin T., Maracci C., Graf M., Karki P., Klepacki D., Berninghausen O., Beckmann R., Vázquez-Laslop N., Wilson D.N., Rodnina M.V, & Mankin A.S. (2017). An antimicrobial peptide that inhibits translation by trapping release factors on the ribosome. Nature structural & molecular biology, 24(9), 752-757.
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3

Rapid Kinetics of Ribosome Release

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Rapid kinetics measurements were performed on an SX-20MV stopped-flow apparatus (Applied Photophysics, Leatherhead, UK). Experiments were performed by rapidly mixing equal volumes (60 μl) of f[3H]Met-tRNAfMet(Flu)-carrying PreHC (0.05 μM), preincubated with Api137 for 2 min at room temperature and RF1Qsy (0.15 μM) at 37°C. Fluorescein was excited at 470 nm and fluorescence emission was monitored after passing a KV500 filter (Schott). Time courses were evaluated by fitting using exponential functions by GraphPad Prism software. Dissociation rates (koff) were determined by chase experiments: PreHCflu (0.05 μM) was preincubated with 0.15 μM RF1Qsy to generate PostHCflu in the absence or presence of 1 μM Api137. PreHC was then rapidly mixed with a 10-fold excess of unlabeled RF1 and RF3·GTP (1 mM); pyruvate kinase (0.1 mg/ml), and phosphoenol pyruvate (3 mM) were present in both syringes. The increase of fluorescence upon dissociation of RF1Qsy was monitored as described above.
Florin T., Maracci C., Graf M., Karki P., Klepacki D., Berninghausen O., Beckmann R., Vázquez-Laslop N., Wilson D.N., Rodnina M.V, & Mankin A.S. (2017). An antimicrobial peptide that inhibits translation by trapping release factors on the ribosome. Nature structural & molecular biology, 24(9), 752-757.
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