CERTIFICATE OF ANALYSIS

PageRuler

Prestained Protein Ladder 

#65-0671   2x250ul Ready-Load

(for 100 mini-gel applications 5ul per well or 50 large gel applications 10ul per well)

Lot: 5929   Quality guaranteed: 12.2009

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

1. Thaw the Ladder 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 per well for mini-gels, 3ul per well for blots,

    10ul per well for large gels, 6ul per well for blots.

4. After the run is complete, stain the gel or perform Western  transfer 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.