90 μg of trichloroacetic acid (TCA) precipitated M. ulcerans (NM20/02) protein lysate was resuspended in rehydration buffer (8 M urea, 2% 3-[(3-Cholamidopropyl)-dimethylammonio]-1-propanesulfonate (CHAPS), 0.5% (v/v) ZOOM Carrier Ampholytes (Invitrogen), 0.002% bromophenol blue and 0.4% dithioerythritol (DTE)). The mix was incubated with a 3–10 pH gradient strip (ZOOM Strip; Invitrogen) over night (ON) at room temperature (RT). First-dimension isoelectric focusing (IEF) was performed on a ZOOM IPG runner (Invitrogen) using a step voltage protocol (175 V for 15 min, 175–2000 V for 45 min, 2000 V for 2 h). After IEF, the strips were incubated for 15 min with equilibration buffer (6 M urea, 50 mM Tris pH 8.8, 30% glycerol, 2% SDS, 30 mM DTE) followed by a 15 min incubation period with alkylating solution (6 M urea, 50 mM Tris (pH 8.8), 30% glycerol, 2% SDS, 0.23 M iodacetamide). Second-dimension gel electrophoresis was performed at 200 V for 35 min using a 10% NuPAGE Novex Bis-Tris ZOOM Gel (Invitrogen). The gel was stained with Coomassie blue (Invitrogen).
Zoom strip
Manufactured by Thermo Fisher Scientific
The ZOOM Strip is a versatile laboratory instrument designed for the rapid and accurate measurement of various parameters in liquid samples. It combines a compact, user-friendly design with advanced sensor technology to provide reliable and consistent results. The ZOOM Strip is suitable for a wide range of applications in research, industrial, and clinical settings.
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Lab products found in correlation
Coomassie blue, by Thermo Fisher Scientific (1 mentions)
Nupage novex bis tris zoom gel, by Thermo Fisher Scientific (1 mentions)
2 d clean up kit, by GE Healthcare (1 mentions)
Dithiothreitol (dtt), by Thermo Fisher Scientific (1 mentions)
Iodoacetamide, by Merck Group (1 mentions)
Silverquest silver staining kit, by Thermo Fisher Scientific (1 mentions)
2 protocols using zoom strip
1
Proteomic Analysis of M. ulcerans
2
2D Gel Electrophoresis of Islet Proteins
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Islet protein extracts were separated by standard 2D-gel electrophoresis by using ZOOM IPGRunner first-dimension isoelectric focusing unit (Invitrogen) and followed by seconddimension 12.5% SDS-PAGE. Prior to isoelectric focusing (IEF), proteins were precipitated away from detergents, salts, lipids, and nucleic acids using the 2-D Clean-Up Kit (GE Healthcare, Pittsburgh, PA). Samples were denatured for 60 min at room temperature in sample buffer containing ZOOM 2D protein solubilizer I, 10 mM DTT, and 0.5% (w/v) 3-10 carrier ampholytes (Invitrogen). Rehydration of ZOOM Strip (pH 3-10, non-linear, Invitrogen) was carried out overnight at room temperature. After IEF, strips were alkylated with 125 mM iodoacetamide (Sigma). Second dimension resolution was carried out by standard SDS-PAGE, followed by protein staining or transfer to nitrocellulose for immunoblotting. Silver staining of 2D gels prior to spot excision and mass-spectroscopy was carried out using the SilverQuest silver staining kit (Invitrogen).
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