Handcast gels must be prepared from acrylamide and bisacrylamide monomer solutions; the component solutions are prepared, mixed together, and then poured between two glass plates to polymerize. This section provides general tips for handcasting gels, protocols for handcasting single and gradient gels, and a list of Bio-Rad products available for handcasting gels.
Related Topics: Polyacrylamide Gels, Buffer Systems and Gel Chemistries, and Protein Standards.
Find more information on hand casting single and gradient SDS-PAGE gels in the Protocols section below.
For more about acrylamide polymerization, refer to Bio-Rad bulletin 1156.
Premade Buffers and Reagents Electrophoresis buffers and reagents are available as individual reagents or as premixed gel-casting, sample, and running buffers. Use of commercially prepared, premixed buffers helps save time but also helps to maximize reproducibility, avoid potential mistakes in buffer concentration and standardize electrophoresis runs, as they are made with electrophoresis-purity reagents and are quality controlled for reproducible results. There are no reagents to weigh or filter; just dilute with distilled or deionized water.
AnyGel™ Stands AnyGel stands provide stabilization and access to gels for casting and sample loading. The clamping mechanism secures gels cassettes vertically without excess pressure. They are available in two sizes, single- and six-row.
Multi-Casting Chambers Multi-casting chambers are sued to cast multiple gels of various thicknesses simultaneously. Acrylic blocks act as space fillers when fewer than the maximum number gels are cst. These chambers work in concert with the gradient formers through a bottom filling port to ensure reproducibility. Multi-casting chambers are available for casting gels for the Mini-PROTEAN®, PROTEAN® II, and PROTEAN® Plus systems.
Gradient Formers Gradient gels have a gradient of acrylamide concentration that increases from top to bottom. To create this gradient, the acrylamide solution must be mixed in a gradient former before being introduced into the gel cassette. Typically, two solutions are prepared: the light solution (equivalent to the lowest %T in the range to be poured) and a heavy solution (equivalent to the maximum %T to be poured). The most common gradient gel contains 4–20% acrylamide; however, the range of acrylamide concentrations should be chosen on the basis of the size of the proteins being separated.
Two gradient formers are available for PAGE systems. Depending on the gel format, prepare either a single gel using the gradient former, or couple the gradient former with a multi-casting chamber for the preparation of up to 12 gels simultaneously:
Multi-casting chambers and gradient formers.
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