Total, nuclear or cytoplasmic protein extracts, or IP samples were prepared in NuPAGE 4X loading buffer (Life Technologies) and 10X reducing agent and hit at 96°C for 5 min. Separations of proteins were performed using pre-cast polyacrylamide gel cassettes (NuPAGE® Novex® 4-12% Bis-Tris) (Life technologies) and 1X NuPAGE MOPS SDS Running Buffer and transferred into nitrocellulose membranes in 20 mM phosphate transfer buffer (pH 6.7). Membranes were blocked in PBS supplemented with 0.1% Tween 20 and 5% non-fat dried milk and incubated with primary antibodies. Primary antibodies were detected with IRDYE- conjugated secondary antibodies and scanned on the LI-COR imaging system. Quantification of bands was performed with LI-COR software.
Nupage 4x loading buffer
Manufactured by Thermo Fisher Scientific
NuPAGE 4X loading buffer is a concentrated solution used to prepare samples for electrophoresis on NuPAGE pre-cast gels. It is designed to ensure proper sample loading and migration on the gel.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using nupage 4x loading buffer
1
Western Blot Analysis of Protein Extracts
2
Western Blot Protein Analysis Protocol
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Total, nuclear or cytoplasmic protein extracts, or IP samples were prepared in NuPAGE 4X loading buffer (Life Technologies) and 10X reducing agent and hit at 96°C for 5 min. Separations of proteins were performed using pre-cast polyacrylamide gel cassettes (NuPAGE® Novex® 4-12% Bis-Tris) (Life technologies) and 1X NuPAGE MOPS SDS Running Buffer and transferred into nitrocellulose membranes in 20 mM phosphate transfer buffer (pH 6.7). Membranes were blocked in PBS supplemented with 0.1% Tween 20 and 5% nonfat dried milk and incubated with primary antibodies.
Primary antibodies were detected with IRDYE-conjugated secondary antibodies and scanned on the LI-COR imaging system. Quantification of bands was performed with LI-COR software.
Primary antibodies were detected with IRDYE-conjugated secondary antibodies and scanned on the LI-COR imaging system. Quantification of bands was performed with LI-COR software.
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