Proteins were separated using 4–12% gradient polyacrylamide SDS–PAGE gels (Life Technologies) and electrotransferred to 0.2 μM nitrocellulose membranes (GE Healthcare, Velizy-Villacoublay, France). After blocking in Tris-buffered saline with 0.1% Tween and 5% bovine serum albumin, membranes were blotted overnight at 4 °C with the appropriate primary antibodies. Primary antibodies were detected using the appropriate horseradish peroxidase-conjugated secondary antibodies. Immunoreactive bands were visualized by enhanced chemiluminescence (PI32209; Thermo Fisher Scientific, Bremen, Germany) with a Syngene camera. Densitometric analyses of immunoblots were performed using the GeneTools software.
Gradient polyacrylamide sds page gels
Manufactured by Thermo Fisher Scientific
Sourced in France, United States
4–12% gradient polyacrylamide SDS–PAGE gels are used for the separation and analysis of proteins based on their molecular weight. These gels provide a range of pore sizes for effective separation of a wide variety of protein samples.
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3 protocols using gradient polyacrylamide sds page gels
1
Immunoblot Analysis of Protein Samples
2
Western Blot Protein Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Proteins were separated using 4–12% gradient polyacrylamide SDS–PAGE gels (Life Technologies) and electrotransferred to 0.2 µm nitrocellulose membranes (GE Healthcare). After blocking in Tris-buffered saline with 0.1% Tween and 5% bovine serum albumin, membranes were blotted overnight at 4 °C with the appropriate primary antibodies. Primary antibodies were detected using the appropriate horseradish peroxidase-conjugated secondary antibodies. Immunoreactive bands were visualized by enhanced chemiluminescence (PI32209; Thermo Fisher Scientific) with a Syngene camera. Densitometric analyses of immunoblots were performed using the GeneTools software. All full scans of uncropped blots are available in the Supplementary file (Supplementary Fig. 8 ).
3
Western Blot Protein Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Proteins were separated using 4–12% gradient poly-acrylamide SDS-PAGE gels (Life Technologies) and electrotransferred to 0.2 µm nitrocellulose membranes (GE Healthcare, Chicago, IL, USA). After blocking with Tris-Buffered Saline with 0.1% Tween and 3% bovine serum albumin, membranes were incubated overnight at 4 °C under continuous shaking with the appropriate primary antibodies. Primary antibodies were detected using the appropriate secondary antibodies coupled with horseradish peroxidase. Immunoreactive bands were visualized by enhanced chemiluminescence with a Syngene camera. Densitometric analyses of immunoblots were performed using the GeneTools software.
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