Description
CERTIFICATE OF ANALYSIS
PageRuler
Prestained Protein Ladder
#65-0671 250ul Ready-Load
(for 100 mini-gel applications 5ul per well or 50 large gel applications 10ul per well)
Store at -20℃
Description
PageRuler Prestained Protein Ladder is a mixture of 10 recombinant, highly purified colored proteins with the apparent molecular weights from 10kDa to 170kDa. It is designed for monitoring of protein separation during SDS-PAGE, verifying Western transfer efficiency and approximating the molecular weight of blotted proteins (1-3). Proteins are covalently coupled with a blue chromophore except for a reference one- 72kDa, coupled with an orange dye. The PageRuler Prestained Protein Ladder is supplied in gel loading buffer and is ready-to-use.
Contents
Approximately 0.1-0.2mg/ml of each protein in the storage buffer (62.5mM Tris-H3PO4 (pH 7.5 at 25), 1mM EDTA, 2% SDS, 10mM DTT, 1mM NaN3 and 33% glycerol).
Applications
- Monitoring of protein separation during SDS-PAGE (1).
- Verifying Western transfer efficiency (2,3).
- Approximating the molecular weight of blotted proteins.
Instruction for Use
- Thaw the Ladder at room temperature or heat at 37-40℃ for a few minutes to dissolve precipitated solids.DO NOT BOIL!
- Mix gently, but thoroughly to ensure the solution is homogeneous.
- Load the Ladder on an SDS-PAA gel and run.
Load volume.
5ul per well for mini-gels, 3ul per well for blots,
10ul per well for large gels, 6ul per well for blots.
- After the run is complete, stain the gel or perform Westerntransfer procedure as desired.
Note
- Each lot of the PageRuler Prestained Protein Ladder is calivrated against a precisely sized, unstained PageRuler Protein Ladder and calculated apparent molecular weights are reported in the picture.
- For precise molecular weight determinations use unstained PageRuler Protein Ladder, #65-0661.
- 8-10% gels may cause protein with low molecular weights to migrate with the dye front.
- Loading volumes are intended for use in gels with a thickness of 0.75mm. If thicker gels are used, the loading volume should be increased.
- PageRuler Prestained Protein Ladder could be used in Western blotting with all common membranes: PVDF, nylon and nitrocellulose.
- Longer transfer times or higher voltages may be required for Western blotting of large (>100kDa) proteins.