(a) Transfer the desired amount of the Prestained Protein Marker to a separate tube. For visualizing during electrophoresis: use 15 µl for mini-gels and 30 µl for full length gels.
(b) Heat the Marker to 95-100ºC for 3-5 minutes.
(c) After a quick microcentrifuge spin, load directly onto a gel. To ensure uniform mobility, load an equal volume of 1X reducing SDS Loading Buffer into any unused wells.
Note on Apparent Molecular Weights:
The relative sizes of these protein markers may depend on the type of gel used and may appear different than expected. The coupling of a charged dye molecule to a protein marker alters the overall charge of the protein and will likely alter its mobility in an SDS polyacrylamide gel. The extent of this effect can vary with the properties of the gel type (e.g., Tris-glycine, Tris-Tricine, etc) used in the analysis. For this reason, the sizes of these marker proteins are expressed here as apparent molecular weights. For best results, we recommend using these prestained protein markers on a Tris-glycine SDS gel.
Contains 0.1–0.2 mg/ml of each protein
Store at -20°C.
|Protein||Source||Apparent MW (Da)|
|CBD-Mxe Intein-2CBD||E. coli||46,000|
|CBD-Mxe Intein||E. coli||30,000|
|Lysozyme||chicken egg white||17,000|
Prestained Protein Marker, Broad Range is a mixture of purified proteins covalently coupled to a blue dye that resolves to 8 bands between 7 and 175 kDa when electrophoresed (1). The protein concentrations are carefully balanced for even intensity. The covalent coupling of the dye to the proteins affects their electrophoretic behavior in SDS-PAGE gels relative to unstained proteins. The apparent molecular weight of the prestained proteins are given in the table below.
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