Description
CERTIFICATE OF ANALYSIS
Protein Ladder, 10-200kDa
2×0.25ml
Lot: 0201 Quality guaranteed: 05. 2006
Store at -20℃
Description
Protein Ladder, 10-200kDa, is a mixture of 14 recombinant, highly purified proteins designed for precise sizing of proteins by SDS-PAGE. The proteins resolve into clearly identifiable sharp bands from 10kDa to 200kDa when analyzed by SDS-PAGE and stained with Coomassie Blue (1). The Protein Ladder is supplied in gel loading buffer and is ready-to-use. Each protein contains an integral Strep-tag® II sequence and may be detected on Western blots using Strep-Tactin® -AP conjugate or antibody against Strep-tag® II (2,3).
Contents
Approximately 0.02-0.05mg/ml of each protein in the following storage buffer (62.5mM Tris-H3PO4 (pH 7.5 at 25), 1mM EDTA, 2% SDS, 10mM DTT, 1mM NaN3, 0.01% bromophenol blue and 33% glycerol).
Application
Precise molecular weight determination of polypeptides in denaturing PAA gels and Western blots (1, 4).
Instruction for Use
1. Thaw the marker at room temperature or heat at 37-40℃ for a few minutes to dissolve precipitated solids. DO NOT BOIL!
2. Mix gently, but thoroughly to ensure the solution is homogeneous.
3. Load the Ladder on an SDS-PAA gel and run.
Load volume.
5ul for mini-gels, 3ul for blots,
10ul for large gels, 6ul for blots.
4. After the run is complete, stain the gel or perform
Western transfer procedure as desired.
Note
The 50k Da protein band is of greater intensity and serves as a reference.
Loading volumes are indicated for in gels with a thickness of 0.75mm. If thicker gels are used, the loading volume should be increased.
When Western blots are indicated for in gels with a thickness of 0.75mm. If thicker gels are used, the loading volume should be increased.
When Western blots are performed, Prestained Protein Ladder (#65-0671) might be used for monitoring of transfer efficiency.
The 200kDa band has lower intensity than the other bands when detected on Western blots using Strep-Tactin® -AP conjugate.
The Protein Ladder is optimized for use with Coomassie Brilliant Blue R-250 staining. For silver staining the amount of Protein Ladder applied might be decreased up to 10 times.
