What is a phos-tag gel?

What is a phos-tag gel?

Phos-tag gels are recent tools to dissect protein phosphorylation that operate by inducing a shift in the electrophoretic mobility of phosphorylated proteins compared to their nonphosphorylated counterparts.

Does the stacking gel solution contains acrylamide?

The upper or stacking gel contains 4-5% acrylamide (a very loose gel) weakly buffered at pH 9.0. The lower resolving gel (often called the running gel), contains a higher acrylamide concentration, or a gradient of acrylamide, strongly buffered at pH 9.0.

What will happen to the gel if the acrylamide concentration is increased?

Increased concentrations of acrylamide result in decreased pore size after polymerization. Polyacrylamide gel with small pores helps to examine smaller molecules better since the small molecules can enter the pores and travel through the gel while large molecules get trapped at the pore openings.

What is the of acrylamide used in resolving gel in page?

Resolving gel: 12.5%: 12.5% acrylamide (A), 0.1% bisacrylamide(B); 15%: 15% A, 0.087% B; 20%: 20% A, 0.066% B.

What is the principle of a phos tag gel and what can be achieved with it?

Phos-tag SDS-PAGE is a phosphate-affinity electrophoresis technique that separates phosphorylated and non-phosphorylated proteins using conventional SDS-PAGE procedures. This technique uses a separation gel in which acrylamide is co-polymerized with Phos-tag Acrylamide that acts as a phosphate-trapping molecule.

What is the purpose of BIS-acrylamide?

Bis-acrylamide is used to create crosslinks between acrylamide to generate polyacrylamide gel in electrophoresis gels. The ratio of bis-acrylamide to acrylamide manipulates the porous characteristics of the polyacrylamide gel.

What is the purpose of acrylamide and bis acrylamide?

Acrylamide: Acrylamide is used as a monomer for polyacrylamide, and used as a binding and thickening agent. Bisacrylamide: Bisacrylamide is mainly used as a crosslinking agent in polymerization processes.

What is Phos tag page?

Phos-tag SDS-PAGE is a phosphate-affinity electrophoresis technique that separates phosphorylated and non-phosphorylated proteins using conventional SDS-PAGE procedures. 1, 2) This technique uses a separation gel in which acrylamide is co-polymerized with Phos-tag Acrylamide that acts as a phosphate-trapping molecule.