Whatman anotop 25

Manufactured by GE Healthcare

Sourced in United Kingdom

The Whatman Anotop™25 is a syringe filter designed for the filtration of samples prior to analysis. It features a 25 mm diameter polycarbonate membrane with a 0.02 μm pore size, suitable for the removal of particulates and microorganisms from liquid samples.

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Lab products found in correlation

Ns500, by Malvern Panalytical (3 mentions) Nta 3, by Malvern Panalytical (2 mentions) Nanosight ns500, by Malvern Panalytical (1 mentions)

5 protocols using whatman anotop 25

EV Characterization using NS500 Instrument

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The EV samples were vortexed and diluted in PBS (0.02‐μm‐filtered; Whatman Anotop™25; GE Healthcare Life Science, Buckinghamshire, UK) to be within the recommended concentration (1.0–9.0 × 10⁸ particles per mL). The samples were loaded into the NS500 instrument (Malvern, Amesbury, UK) by a syringe at a constant flow with a syringe pump speed of 20. Three 60‐s videos were captured for each sample (slide shutter 1200, slider gain 146). The videos were analyzed by nta 3.1 software (Malvern). The vesicle quantifications had an analytic variance of 2–25% [17].

Bjørnetrø T., Steffensen L.A., Vestad B., Brusletto B.S., Olstad O.K., Trøseid A., Aass H.C., Haug K.B., Llorente A., Bøe S.O., Lång A., Samiappan R., Redalen K.R., Øvstebø R, & Ree A.H. (2021). Uptake of circulating extracellular vesicles from rectal cancer patients and differential responses by human monocyte cultures. FEBS Open Bio, 11(3), 724-740.

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Feraheme Removal via Filtration

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In order to determine whether Feraheme could be easily removed from solution, a simple filtration was attempted. Feraheme doped solutions were forced through a Whatman Anotop 25 syringe filter with 0.02 μm pores (GE Healthcare, UK). This was performed both on a mock dissolution sample of water with Feraheme and on a sample that had been polarized and dissolved. The mock sample contained 0.268 mM elemental iron in deionized water to mimic the concentration of Feraheme in a post-dissolution DNP sample. This sample was evaluated by measuring UV-Vis absorbance and proton T₁ before and after filtration. The ¹³C T₁ of the dissolution sample was measured pre-filtration by the decay of the hyperpolarized signal and measured post-filtration by inversion recovery in the same magnet.

Niedbalski P., Parish C.R., Wang Q., Hayati Z., Song L., Cleveland Z.I, & Lumata L. (2017). Enhanced Efficiency of 13C Dynamic Nuclear Polarization by Superparamagnetic Iron Oxide Nanoparticle Doping. The journal of physical chemistry. C, Nanomaterials and interfaces, 121(35), 19505-19511.

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Nanoparticle Tracking Analysis of Extracellular Vesicles

Cited 1 time

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EV isolates were analyzed by nanoparticle tracking analysis as previously described^{56 (link)}. Briefly, EVs were diluted in 0.02 µm-filtered PBS (Whatman Anotop™25, GE Healthcare Life Science, Buckinghamshire, UK), to reach the measurement range (1.0–9.0 × 10⁸ particles/mL). Samples were loaded into a NanoSight NS500 instrument (Malvern, Amesbury, UK) equipped with a sCMOS camera, using a syringe pump and flow speed 20. For each sample, three 60-s videos were captured, typically at camera level 14 and detection threshold 3, then analyzed using NTA 3.1 software Build 3.1.54.

Vestad B., Nyman T.A., Hove-Skovsgaard M., Stensland M., Hoel H., Trøseid A.M., Aspelin T., Aass H.C., Puhka M., Hov J.R., Nielsen S.D., Øvstebø R, & Trøseid M. (2021). Plasma extracellular vesicles in people living with HIV and type 2 diabetes are related to microbial translocation and cardiovascular risk. Scientific Reports, 11, 21936.

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Tear Fluid EV Isolation and Characterization

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EVs were isolated from a pooled tear fluid sample using size-exclusion chromatography (SEC) (qEV 70 nm size-exclusion chromatography column, iZON Science, Oxford, UK). Fractions 7–10 were combined, and the size distribution and concentration of the EVs were determined by nanoparticle tracking analysis (NTA), using a NanoSight NS500 (Malvern Instruments Ltd., Amesbury, UK). The sample was diluted 1:1 with filtered PBS (0.02 μm filter, Whatman™ Anotop™ 25, GE Healthcare Life Science, Buckinghamshire, UK), vortexed, and then analyzed in triplicate. The camera level was set to 13, and the detection threshold was set to 3. Videos 60 s in length of a continuous flow of sample were recorded. Data analysis was performed using NTA 3.1 software.

Cross T., Øvstebø R., Brusletto B.S., Trøseid A.M., Olstad O.K., Aspelin T., Jackson C.J., Chen X., Utheim T.P, & Haug K.B. (2023). RNA Profiles of Tear Fluid Extracellular Vesicles in Patients with Dry Eye-Related Symptoms. International Journal of Molecular Sciences, 24(20), 15390.

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Extracellular Vesicle Size and Concentration

Cited 1 time

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The EVs were analyzed with a nanoparticle tracking analysis instrument (NS500, Malvern, Amesbury, United Kingdom) to determine the vesicle concentration and size distribution. The EV samples were diluted by PBS (0.02 μm-filtered; Whatman Anotop™25, GE Healthcare Life Science, Buckinghamshire, United Kingdom) to be within the recommended concentration range (1.0-9.0×10⁸ particles/mL). Samples were loaded into the NS500 instrument at a constant flow with a syringe pump speed of 20. Triplicates of 60-s videos were captured for each sample. The videos were analyzed by NTA 3.1 software (Malvern). The vesicle quantifications, as previously described, had an intra-assay coefficient of variance (CV) below 7% and inter-assay CV (day-to-day) < 20% (Vestad et al., 2017 (link)).

Aas V., Øvstebø R., Brusletto B.S., Aspelin T., Trøseid A.M., Qureshi S., Eid D.S., Olstad O.K., Nyman T.A, & Haug K.B. (2023). Distinct microRNA and protein profiles of extracellular vesicles secreted from myotubes from morbidly obese donors with type 2 diabetes in response to electrical pulse stimulation. Frontiers in Physiology, 14, 1143966.

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