SDS-PAGE Analysis of rFVIII

Ready Gel^® Tris-HCl Gels (4–15%) (Bio-Rad, USA) were run according to standard protocol [20 (link)]. The rpFVIII sample at 2 µg/well concentration was reduced with 5% β-mercaptoethanol in Bio-Rad Laemmli Sample buffer at 94°C for 5 min, loaded onto gels with Bio-Rad Pre-stained Standards Broad Range and run at 115 V for 2 hours 30 minutes. The gels were stained with GelCode^® Blue Stain Reagent (Thermo Scientific, USA) and de-stained in water.

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Csencsits-Smith K., Grushin K, & Stoilova-McPhie S. (2015). Binding of Factor VIII to Lipid Nanodiscs Increases its Clotting Function in a Mouse Model of Hemophilia A. Journal of blood disorders & transfusion, 6(6), 325.

Publication 2015

Ready Gel® Tris-HCl Gels (4–15%) Laemmli Sample buffer GelCode® Blue Stain Reagent

Gels Laemmli buffer Mercaptoethanol Tris

Corresponding Organization :

Other organizations : The University of Texas Health Science Center at Houston, The University of Texas Medical Branch at Galveston

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Variable analysis

independent variables

Reducing agent (5% β-mercaptoethanol)
Incubation temperature (94°C)
Incubation time (5 min)

dependent variables

Mobility of rpFVIII sample

control variables

Gel type (Ready Gel® Tris-HCl Gels (4–15%))
Gel loading (2 µg/well concentration of rpFVIII sample)
Electrophoresis voltage (115 V)
Electrophoresis duration (2 hours 30 minutes)
Staining (GelCode® Blue Stain Reagent)
Destaining (water)

positive controls

Bio-Rad Pre-stained Standards Broad Range

negative controls

Not explicitly mentioned

Annotations

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