Purified rFIX samples were separated by SDS–PAGE using a 4–12% Bis-Tris Bolt acrylamide gel (Invitrogen) in 1× MES running buffer (Invitrogen), under nonreducing conditions, following manufacturer’s instructions. Protein bands were visualized either with colloidal Coomassie blue (Simply Blue; Invitrogen) or western blotting. For western blotting, proteins were transferred from gels onto a PVDF membrane (Bio-Rad) using a Trans-Blot® Turbo™ Transfer System Transfer Pack (Bio-Rad), following manufacturer’s instructions. The membrane was blocked in 2% skim milk (Devondale) dissolved in phosphate-buffered saline solution (Sigma) with 0.05% Tween 20 (Sigma) for 30 min at room temperature. The following antibodies were used: mouse IgG1 anti human FIX primary antibody 1C2 (AbCam, 1:1000 dilution) and HRP-conjugated anti mouse IgG (Bio-RAD, 1:1000 dilution). Membranes were developed with Novex ECL reagent (Invitrogen) and imaged using a Bio-Rad image workstation.
Bis tris bolt acrylamide gel
Manufactured by Thermo Fisher Scientific
The 4–12% Bis-Tris Bolt acrylamide gel is a pre-cast electrophoresis gel designed for the separation and analysis of proteins. It features a gradient of 4% to 12% polyacrylamide concentration, which allows for the effective resolution of a wide range of protein sizes. The gel is formulated with Bis-Tris buffer, which provides superior pH control and protein separation performance.
Automatically generated - may contain errors
Lab products found in correlation
Image workstation, by Bio-Rad (2 mentions)
Novex ecl reagent, by Thermo Fisher Scientific (2 mentions)
Simplyblue, by Thermo Fisher Scientific (2 mentions)
Tween 20, by Merck Group (2 mentions)
Pvdf membrane, by Bio-Rad (2 mentions)
Mes running buffer, by Thermo Fisher Scientific (2 mentions)
Phosphate buffered saline solution, by Merck Group (1 mentions)
Trans blot turbo transfer system transfer pack, by Bio-Rad (1 mentions)
Phosphate buffered saline solution pbs, by Merck Group (1 mentions)
2 protocols using bis tris bolt acrylamide gel
1
SDS-PAGE and Western Blot Analysis of rFIX
2
Purified rFIX Protein Analysis
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Purified rFIX samples were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) using a 4-12% Bis-Tris Bolt acrylamide gel (Invitrogen) in 1x MES running buffer (Invitrogen), under non-reducing conditions, following manufacturer's instructions. Protein bands were visualized either with colloidal Coomassie blue (Simply Blue; Invitrogen) or western blotting. For western blotting, proteins were transferred from gels onto a PVDF membrane (Bio-Rad) using a Trans-Blot ® Turboä Transfer System Transfer Pack (Bio-Rad), following manufacturer's instructions. The membrane was blocked in 2% skim milk (Devondale) dissolved in phosphate buffered saline solution (PBS; Sigma) with 0.05% Tween 20 (Sigma) for 30 min at room temperature. The following antibodies were used: mouse IgG1 anti human Factor IX primary antibody 1C2 (AbCam, 1:1000 dilution) and HRP-conjugated anti mouse IgG (BioRAD, 1:1000 dilution). Membranes were developed with Novex ECL reagent (Invitrogen) and imaged using a Bio-Rad image workstation.
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