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Protein Prestained Protein Ladder V..................................................................81 Prestained Protein Ladder 245 kDa.......................................................81 Protein Loading D ye..............................................................................82 R everse Protein Stain K it.......................................................................82 Protein Prestained Protein Ladder V Introduction The Prestained Protein Ladder V is a three-color prestained protein standard which includes protein dye for convenient protein sample staining. The protein ladder consists of 10 prestained proteins covering a wide range of molecular weights (10 to 180 kDa). Proteins are covalently coupled with a blue chromophore except for two reference bands (one green and one red band) when separated on SDS-PAGE. The ladder is supplied in a gel loading buffer and is ready to use, without heating, diluting, or a reducing agent prior to loading. Advantages • Wide Molecular Weight range: 10-180 kDa • Supplied buffer for direct gel loading • Sharp bands with 28 and 75 KDa reference bands (green/red dye) • Includes Protein Loading Dye (2 ml, 5X) • 3 μl or 5 μl per loading for clear visualization during electrophoresis on 15-well or 10-well mini-gel, respectively • 2~3 μl per well for general Western transferring • Storage: 25ºC for 2 weeks, 4ºC for 3 months, -20ºC for 24 months Prestained Protein Ladder V Molecular Weight Estimation Applications Monitor protein migration/sample in SDS-PAGE, Monitor protein transfer onto membranes during Western Blotting, Sizing of proteins on SDS-PAGE and Western blots Components Approximately 0.2~0.4 mg/ml of each protein in buffer {20 mM Trisphosphate pH7.5 at 25°C, 2% SDS, 1 mM 2-Mercaptoethanol, 3.6 M Urea and 15% (v/v) glycerol}, Protein Loading Dye (2 ml, 5X) Figure 1. Migration patterns of various electrophoretic conditions according to type and percentage of running buffer. Prestained Protein Ladder 245 kDa Introduction Prestained Protein Ladder 245 kDa Molecular Weight Estimation Prestained Protein Ladder (245 kDa) is a three-color protein standard with 12 pre-stained proteins covering a wide range of molecular weights for 10 to 245 kDa. Proteins are covalently coupled with a blue chromophore except for two reference bands (one green and one red band) when separated on SDS-PAGE (Tris-glycine buffer). The ladder is supplied in a gel loading buffer and is ready to use, without requiring heating, diluting, or a reducing agent prior to loading. Advantages • Wide Molecular Weight range: 10-245 kDa • Supplied buffer for direct gel loading • Sharp bands with 28 and 75 KDa reference bands (green/red dye) • Includes Protein Loading Dye (2 ml, 5X) • 3 μl or 5 μl per loading for clear visualization during electrophoresis on 15-well or 10-well mini-gel, respectively • 2~3 μl per well for general Western transferring • Storage: 25ºC for 2 weeks, 4ºC for 3 months, -20ºC for 24 months Applications Monitor protein migration/sample in SDS-PAGE, protein transfer onto membranes during Western Blotting, sizing of proteins on SDS-PAGE and Western blots Figure 2. Migration patterns of various electrophoretic conditions according to type and percentage of running buffer. Components Approximately 0.2~0.4 mg/ml of each protein in buffer {20 mM Trisphosphate pH7.5 at 25°C, 2% SDS, 1 mM 2-Mercaptoethanol, 3.6 M Urea and 15% (v/v) glycerol}, Protein Loading Dye (2 ml, 5X) Page 81 www.geneaid.com Protein Protein Protein Loading Dye Introduction Geneaid's ready-to-use (5X) Protein Loading Dye is ideal for preparing protein samples to be separated in SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and allows for easy sample monitoring during electrophoresis. Protein Loading Dye is conveniently included with Geneaid's Prestained Protein Ladder V. Advantages • Conveniently included with Geneaid's Prestained Protein Ladder V • Ready-to-use solution • Ideal for preparing protein samples to be separated in SDS-polyacrylamide gel electrophoresis (SDS-PAGE) • Allows for easy sample monitoring during electrophoresis • Storage: 25ºC for 2 weeks, 4ºC for 3 months, -20ºC for 24 months Applications Preparing of protein samples to be separated in SDS-polyacrylamide gel electrophoresis (SDS-PAGE), Easy sample monitoring during electrophoresis Reverse Protein Stain Kit Introduction The Reverse Protein Stain Kit uses imidazole and zinc salts for protein detection as low as 1 ng in electrophoresis gels. The method is based on selective precipitation of a white imidazole–zinc complex in the gel, except in zones where proteins are located which remain transparent. When the gel is placed on a dark background, the negative gel image can be converted to a positive image with black bands, spots, and a white background. This stain is as sensitive as most silver stains and requires only 10 minutes to complete. Fixation of proteins to the gel is not needed, there is no interaction of the stain with the protein, and complete destaining of the matrix can be achieved. Imidazole-zinc reverse stain uses only two staining solutions, which can be easily diluted from stock solutions. For the imidazole-zinc negative staining, no fixation is necessary. Gels are typically stained immediately after electrophoresis. Advantages • Protein detection as low as 1 ng in electrophoresis gels • Fixation of proteins to the gel is not needed so will not affect protein samples • Complete destaining of the matrix can be achieved • Rapid Staining: <10 minutes compared to conventional staining methods requiring up to 24 hours • Non toxic (no organic solvents) • Alternative staining methods can be used direclty following this procedure • Storage: room temperature for up to 2 years Applications Imidazole-zinc reverse stain is fully compatible to down-stream applications, such as Mass spectrometry, Edman sequencing, Electroelution, and Membrane blotting techniques Reverse Protein Stain Kit <10 Minute Gel Figure 1. The Reverse Protein Stain Kit uses imidazole and zinc salts for protein detection as low as 1 ng in electrophoresis gels. The method is based on selective precipitation of a white imidazole–zinc complex in the gel, except in those zones where proteins are located. Components • Protein Stain Buffer I and II • Black Stain Box Protein www.geneaid.com Page 82