Slide a lyzer 10k mwco

Manufactured by Thermo Fisher Scientific

The Slide-A-Lyzer 10K MWCO is a dialysis device designed for the purification and concentration of macromolecules, such as proteins and peptides, with a molecular weight cut-off of 10,000 Daltons. It enables the removal of small molecules from sample solutions through the process of dialysis.

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Lab products found in correlation

Series s sensor chip sa, by GE Healthcare (1 mentions) Biacore t200, by Cytiva (1 mentions) Flaming brown micropipette puller, by Sutter Instruments (1 mentions) Omnisolv lc ms grade, by Merck Group (1 mentions) Superblock pbs blocking buffer, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

Slide-A-Lyzer (133 citations) 10K MWCO (8 citations) 10K MWCO Slide-A-Lyzer Dialysis Cassettes (6 citations) Slide-A-Lyzer Dialysis cassette 10K MWCO (3 citations) 10K MWCO Slide-A-Lyzer G2 Dialysis Cassette (2 citations) 10K MWCO Slide-A-Lyzer cassettes (2 citations)

4 protocols using slide a lyzer 10k mwco

Extraction and Purification of Exopolysaccharide

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The EPS was
isolated and purified following the method of Adesulu-Dahunsi et al.^{11 (link)} with slight modifications. After cell cultivation
and collection, SMW was heated at 100 °C for 10 min to inactivate
the enzymes, and the debris was removed by centrifugation (10 000g for 20 min at 4 °C). Three volumes of 95% (v/v) cold
ethanol were added to the supernatant and stored at −4 °C
overnight and then centrifuged (10 000g for
20 min at 4 °C). Crude EPS (cEPS) was obtained after ethanol
precipitation and lyophilization. To remove the protein, cEPS was
dissolved in distilled water and deproteinized using 4% (v/v) trichloroacetic
acid (TCA). cEPS was reprecipitated with three volumes of 95% (v/v)
cold ethanol and dissolved in distilled water. Finally, dialysis was
conducted twice daily with distilled water for 2 days at 4 °C
using dialysis cassettes (Slide-A-Lyzer 10K MWCO, Thermo Scientific,
Waltham, MA) and then lyophilized. The final residual was termed pure
EPS (pEPS).

Choi I.S., Ko S.H., Lee M.E., Kim H.M., Yang J.E., Jeong S.G., Lee K.H., Chang J.Y., Kim J.C, & Park H.W. (2021). Production, Characterization, and Antioxidant Activities of an Exopolysaccharide Extracted from Spent Media Wastewater after Leuconostoc mesenteroides WiKim32 Fermentation. ACS Omega, 6(12), 8171-8178.

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Investigating Polysaccharide-RNA Interactions

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We analyzed the direct interaction between polysaccharides and RNA using a BIAcore T200 surface plasmon resonance analyzer (GE Healthcare). To prepare ligands, biotinylated 19 R was diluted with HBS-EP buffer, composed of 10 mM HEPES (pH 7.4), 150 mM NaCl, 3 mM EDTA, and 0.05% (v/v) surfactant P, and immobilized on a streptavidin-coated Series S Sensor Chip SA (GE Healthcare) at a flow rate of 10 µL/min for 60 s. All the datasets were collected at 25 °C.
Two-hundred microliters of purified polysaccharides were dialyzed using Slide-A-Lyzer 10 K MWCO (Thermo Fisher Scientific) in 500 mL of HBS-EP overnight at 4 °C. The dialyzed polysaccharides were diluted in HBS-EP buffer and subjected to analytes at a flow rate of 30 µL/min for 30 s in manual mode. Because the molecular-size distribution of polysaccharides purified from the biofilm matrix was vast, the molar concentration of the polysaccharides was difficult to determine. Dispersin B was diluted in HBS-EP to a final concentration of 16 µg/mL and used to digest the bound polysaccharides at a flow rate of 30 µL/min for 30 s.

Chiba A., Seki M., Suzuki Y., Kinjo Y., Mizunoe Y, & Sugimoto S. (2022). Staphylococcus aureus utilizes environmental RNA as a building material in specific polysaccharide-dependent biofilms. NPJ Biofilms and Microbiomes, 8, 17.

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Intact Protein IMS-MS Analysis of Hemoglobin

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For intact protein IMS-MS experiments, human hemoglobin A (Sigma-Aldrich, St. Louis, MO) was prepared in water (Omnisolv LC-MS grade, EMD Millipore, Billerica, MA), and desalted by overnight dialysis (Slide-A-Lyzer 10K MWCO, Thermo Scientific, Rockford, IL) against 150 mM ammonium acetate (pH 7.4). For MS analysis, samples were diluted to 15 μM (tetramer concentration) in 150 mM ammonium acetate (pH 7.4). Fifteen μM protein concentration was chosen for optimal signal-to-noise ratio Analyte solution was loaded into a borosilicate glass nano ESI (nESI) capillary (Sutter Instrument, Novato, CA) that was pulled to a fine point (~1−5 μm) using a Flaming/Brown micropipette puller (model P-97, Sutter Instrument, Novato, CA). If not otherwise stated, all reagents were purchased from Sigma-Aldrich (St. Louis, MO) in the highest purity available.

Woodall D.W., Brown C.J., Raab S.A., El-Baba T.J., Laganowsky A., Russell D.H, & Clemmer D.E. (2020). Melting of Hemoglobin in Native Solutions as measured by IMS-MS. Analytical chemistry, 92(4), 3440-3446.

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Mannose-Purification of Surface Proteins

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The mannose-purification of clarified surface protein extracts of Ac and RAW 264.7 was scaled up to 96-well microplates previously sensitized with 50 µL of mannose solution (10 µg/mL) for 1 h at 37°C, and subsequently overnight at 4°C. After, the plates were blocked with blocking buffer (SuperBlock™ (PBS) Blocking Buffer, ThermoFisher), for 1 h at 37°C. Thereafter, plates were incubated with 200 µg/mL (50 µL/well) of phagocytes’ clarified extracts in DPBS, for 1 h at 37°C. Posteriorly, plates were washed three times with PBS to remove mannose unbound proteins. Then, mannose-attached proteins were recovered with stripping buffer (50 mM Tris-HCl, 50 mM DTT, and 2% SDS; pH 7.0) for 1 h at 70°C. Next, the content of each well was collected and dialyzed in dialysis cassettes (Slide-A-Lyzer™ 10K MWCO, ThermoFisher) against PBS/Ca⁺²/Mg⁺² at 4°C, under 200 rpm shaking for three days, with daily buffer exchanges. Then, the mannose-purified surface proteins (MPPs) were collected, identified, and analyzed by Western blot. The protein banding pattern of the MPPs were compared between the distinct phagocytes.

Ferreira M.D., Mendoza S.R., Gonçalves D.D., Rodríguez-de la Noval C., Honorato L., Nimrichter L., Ramos L.F., Nogueira F.C., Domont G.B., Peralta J.M, & Guimarães A.J. (2022). Recognition of Cell Wall Mannosylated Components as a Conserved Feature for Fungal Entrance, Adaptation and Survival Within Trophozoites of Acanthamoeba castellanii and Murine Macrophages. Frontiers in Cellular and Infection Microbiology, 12, 858979.

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