The resuspended Euglena cells from the culturing (1 ml) were diluted with 5x binding/wash buffer (0.25 ml) containing phenylmethylsulfonyl fluoride (2 mM) and lysed by sonication (3 × 10 s, 25% amplitude, 30 s off between each pulse) and centrifuged (5 min, 1000 × g). Not all cells were lysed. Total lysate containing the equivalent of 1.1 mg of protein (Easy Bradford BioRad, BSA standards) was then used for glycoprotein purification using both ConA and wheat germ agglutinin (WGA) Glycoprotein Isolation Kits (Thermo Scientific) according to the manufacturer’s instructions. Protein quality was assessed using silver-stained SDS-PAGE (Bolt 4%–12% Bis-TRIS plus, Invitrogen) using SeeBlue Plus2 Prestained Protein Standard (Thermo Fisher Scientific) as the standard.
Easy bradford
Manufactured by Bio-Rad
The Easy Bradford is a colorimetric protein assay kit designed for the quantitative determination of protein concentrations. It utilizes the Bradford method, which is based on the color change of Coomassie Brilliant Blue G-250 dye in response to different protein concentrations. The kit provides a simple, reproducible, and rapid way to measure protein levels in biological samples.
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Lab products found in correlation
Bolt 4 12 bis tris plus, by Thermo Fisher Scientific (2 mentions)
Seeblue plus2 pre stained protein standard, by Thermo Fisher Scientific (2 mentions)
Wga glycoprotein isolation kits, by Thermo Fisher Scientific (2 mentions)
Bsa standard, by Bio-Rad (1 mentions)
Concanavalin a (cona), by Thermo Fisher Scientific (1 mentions)
2 protocols using easy bradford
1
Glycoprotein Isolation and Purification
2
Euglena Glycoprotein Purification Protocol
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The resuspended Euglena cells from the culturing (1 ml) were diluted with 5x Binding/Wash buffer (0.25 ml) containing phenylmethylsulfonyl fluoride (2 mM) and lysed by sonication (3 x 10 s, 25% amplitude, 30 s off between each pulse) and centrifuged (5 min, 1000 xg). Not all cells were lysed. Total lysate containing the equivalent of 1.1 mg of protein (Easy Bradford BioRad, BSA standards) was then used for glycoprotein purification using both ConA and WGA Glycoprotein Isolation Kits (Thermo Scientific) according to the manufacturer's instructions. Protein quality was assed using silver stained SDS-PAGE (Bolt 4-12% Bis-TRIS plus, Invitrogen) using SeeBlue Plus2 Pre-stained Protein Standard (Thermo Fisher Scientific) as the standard.
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