PageRuler™ Unstained Protein Ladder
Lot specific pictures:
18067, 21128, 23980,
28137/6#SM0661 2x250 µl
(for 100 mini gel applications, 5 µl/well
or 50 large gel applications, 10 µl/well)Related Documents (in pdf, ~100 KB):
Certificate of Analysis: #SM0661
MSDS (English)
MSDS (English-USA)
MSDS (German)
LabAid™
Features
- Broad range: 10-200 kDa.
- Ready-to-use – supplied in a loading buffer for direct loading on gels.
- Sharp bands.
- Includes a 50 kDa reference band of a greater intensity.
- Each protein in the ladder contains an integral Strep-tag® II sequence.
Description
The PageRuler™ Unstained Protein Ladder is designed for accurate sizing of proteins by SDS-polyacrylamide gel electrophoresis (1). It is a mixture of 14 recombinant, highly purified, unstained proteins that appear as sharp bands from 10 kDa to 200 kDa on SDS-polyacrylamide gel after staining with PageBlue™ Protein Staining Solution or by other protein staining methods. PageRuler™ Unstained Protein Ladder can also be used in Western blotting on PVDF, nylon and nitrocellulose membranes for precise sizing of blotted proteins.
Each protein in the ladder contains an integral Strep-tag® II sequence which can be detected directly on Western blots (see Western Blotting protocol) using a Strep-Tactin®-AP* conjugate or an antibody against Strep-tag® II (2, 3).
Strep-tag® II detection systems are not supplied by Fermentas.Applications
Accurate protein sizing on SDS-polyacrylamide gels and Western blots.Composition
0.02-0.05 mg/ml of each protein in 62.5 mM Tris-H3PO4 (pH 7.5 at 25°C), 1 mM EDTA, 2% SDS, 100 mM DTT, 1 mM NaN3, 0.01% bromophenol blue and 33% glycerol.
Quality Control
Tested in SDS-polyacrylamide gel electrophoresis and Western blotting.
Storage
Store at -20°C.
Recommendations for Loading
- Thaw the ladder either at room temperature or at 37-40°C for a few minutes to dissolve precipitated solids. Do not boil.
- Mix gently, but thoroughly, to ensure that the solution is homogeneous.
- Load the following volumes of the ladder on SDS-polyacrylamide gel:
- 5 µl per well for mini-gels;
- 10 µl per well for large gels.Note
- The indicated loading volume is recommended for gels with a thickness of 0.75 mm. For thicker gels, the loading volume should be increased.
- A separation gel resolves proteins effectively according to their molecular weight. Linear gradient gels are used for resolution of both small and large proteins, while low percentage gels are recommended for analysis of large proteins. In these gels, small proteins migrate with the tracking dyes during electrophoresis.
Recommendations for Detection
- After the electrophoresis is complete, stain the gel with PageBlue™ Protein Staining Solution, see protocol.
- For silver staining (4), the volume of PageRuler™ Unstained Protein Ladder used should be decreased up to 5-fold.
Related Products
- Laemmli, U.K., Cleavage of structural proteins during the assembly of the head of bacteriophage T4, Nature, 227, 680-685, 1970.
- Schmidt, T.G.M., Skerra, A., The random peptide library-assisted engineering of a C-terminal affinity peptide, useful for the detection and purification of a functional Ig Fv fragment, Prot. Engineering, 6, 109-122, 1993.
- Tsiotis, G., et al., Isolation and structural characterization of trimeric cyanobacterial photosystem I complex with the help of recombinant antibody fragments, Eur. J. Biochem., 231, 823-830, 1995.
- Sorense, B.K., et al., Silver Staining of Proteins on Electro-blotting Membranes and Intensification of Silver Staining of Proteins Separated by Polyacrylamide Gel Electrophoresis, Anal.Biochem., 304, 33-41, 2002.
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Updated vasario 06, 2008 16:37
