Prestained Protein Ladder, 10-170kD is a mixture of 10 recombinant, highly purified colored proteins, designed for monitoring of protein separation during SDS-PAGE, verifying Western transfer efficiency and approximating the molecular weight of blotted proteins. Proteins are covalently coupled with a blue chromophore except two reference one - ~70kD, (orange) and ~10kD (green). The Prestained Protein Ladder is supplied in gel loading buffer and is ready-to-use. Note: 250ul is sufficient for 50 mini-gel applications (5ul/well) or 25 large gel applications (10ul/well) 5ul of Prestained Protein Ladder provide 10 bands of equal intensities in 4-20% SDS-PAGE (Tris-glycine buffer) and after electrotransfer onto PVDF membrane. Applications: Suitable for use in monitoring of protein separation during SDS-PAGE (1), Verifying Western transfer efficiency (2, 3), Approximating the molecular weight of blotted proteins. Protocol: 1. Thaw Ladder at RT. Do not boil! 2. Mix gently, but thoroughly to ensure the solution is homogeneous. 3. Load the Ladder on a gel and run. (Load volume: 5ul for mini-gels, , 10ul for large gels, use same volumes for blots). 4. After run is complete, stain gel or perform Western transfer procedure as desired. Note: <10% gels may cause proteins 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 voume should be increased. Prestained Protein Ladder could be used in Western Blot with all common membranes: PVDF, nylon and nitrocellulose. Longer transfer times or higer voltages may be required for Western Blot of large (>100kD) proteins. Storage and Stability: May be stored at 4C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20C. Aliquots are stable for 6 months at -20C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Formulierung:
Supplied as a liquid in 62.5mM Tris-H3PO4, pH 7.5, 1mM EDTA, 2% SDS, 10mM DTT, 1mM sodium azide, 33% glycerol
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