PageRuler™ Prestained Protein Ladder Plus

Sprawdź także MWP04

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Wcześniejszy numer katalogowy: SM1811

Thermo Scientific PageRuler Plus Prestained Protein Ladder is a mixture of nine (9) blue-, orange- and green-stained proteins (10 to 250kDa) for use as size standards in protein electrophoresis (SDS-PAGE) and Western blotting.

Features of PageRuler Plus Prestained Protein Ladder:

• Size range—nine proteins spanning 10 to 250kDa
• Ready-to-use—supplied in a loading buffer for direct loading on gels; no need to boil
• Sharp bands—color-coded bands of similar intesity for easy visualization
• Quality tested—each lot evaluated by SDS-PAGE and Western blotting
• Bright reference bands—orange at 70 and 25kDa, and green at 10kDa
• Membrane-compatible—colored bands transfer to membranes for Western blotting

This prestained protein MW marker is designed for monitoring the progress of SDS-polyacrylamide gel electrophoresis, for assessing transfer efficiency onto PVDF, nylon and nitrocellulose membranes, and for estimating the approximate size of separated proteins that have been made visible with gel stains or Western blot detection reagents. A blue chromophore is bound to all proteins, except proteins of two reference bands of 70kDa and 25kDa that are colored with an orange dye and one green reference band of 10kDa. PageRuler Plus Prestained Protein Ladder is ready to use: no heating, further dilution or addition of a reducing agent is required before loading onto a gel.

Includes:
• Dye-stained proteins in 62.5 mM Tris-H3PO4 (pH 7.5 at 25°C), 1 mM EDTA, 2% SDS, 10 mM DTT, 1 mM NaN3 and 33% glycerol.

Applications:
• Monitoring protein migration during SDS-polyacrylamide gel electrophoresis
• Monitoring protein transfer onto membranes after Western blotting
• Sizing of proteins on SDS-polyacrylamide gels and Western blots

Note
For migration patterns under different gel conditions, see Specifications.

Recommendations for Loading
1. Thaw the ladder either at room temperature or at 37-40°C for a few minutes to dissolve precipitated solids. Do not boil!
2. Mix gently, but thoroughly, to ensure that the solution is homogeneous.
3. Load the following volumes of the ladder on SDS-polyacrylamide gel:
– 5 μL per well for mini gel,
– 10 μL per well for large gel.
Use the same volumes for Western blotting.
The loading volumes listed above are recommended for gels with a thickness of 0.75-1.0 mm. The loading volume should be doubled for 1.5 mm thick gels.

Important Notes
• Prestained proteins can have different mobilities in various SDS-PAGE-buffer systems. However, they are suitable for approximate molecular weight determination when calibrated against unstained standards in the same system. See the table provided for migration patterns in different electrophoresis conditions.
• In low-percentage gels (< 10%), the low-molecular weight proteins in the ladder may migrate with the dye front.
• PageRuler Plus Prestained Protein Ladder can be used in Western blotting with all common membranes: PVDF, nylon and nitrocellulose.
• Longer transfer times or higher transfer voltages may be required for Western blotting of large (>100 kDa) proteins.