Bio-Rad Prestained SDS-PAGE Standards User Manual

Reference
1. Laemmli, U. K., Nature, 227, 680 (1970).
Ordering Information
Catalog Number Product Description
Prestained Standards
161-0305 Prestained SDS-PAGE Standards, low range, 500µl 161-0309 Prestained SDS-PAGE Standards, high range, 500 µl 161-0318 Prestained SDS-PAGE Standards, broad range, 500 µl 161-0324 Kaleidoscope Prestained Standards, 500 µl 161-0325 Kaleidoscope Polypeptide Standards, 500 µl
Molecular Weight Standards
161-0304 SDS-PAGE Standards, low range, 200 µl 161-0303 SDS-PAGE Standards, high range, 200 µl 161-0317 SDS-PAGE Standards, broad range, 200 µl 161-0314 Silver Stain SDS-PAGE Standards, low range, 200 µl 161-0315 Silver Stain SDS-PAGE Standards, high range, 200 µl 161-0306 Biotinylated SDS-PAGE Standards, low range, 250 µl 161-0311 Biotinylated SDS-PAGE Standards, high range, 250 µl 161-0319 Biotinylated SDS-PAGE Standards, broad range, 250 µl 161-0326 Polypeptide SDS-PAGE Standards, 200 µl
Specialty Standards
161-0310 IEF Standards, pI range 4.45-9.6, 250 µl 161-0320 2-D SDS-PAGE Standards, 500 µl
Bio-Rad Laboratories, 2000 Alfred Nobel Dr., Hercules, CA 94547
4006024 Rev D
Prestained
SDS-PAGE Standards,
Low Range
Catalog Number
161-0305
Product shipped at room temperature.
Store at -20 °C upon arrival
Bio-Rad’s prestained standards are available in high, low, and broad molecular weight ranges. Blue dye has been covalently attached to the standard proteins and will not be dissociated by normal staining or destaining. The protein mixtures are provided in a stable aqueous solu­tion. No reconstitution or further dilution is required before use.
Applications
Prestained SDS-PAGE Standards provide a quick and easy way to assess the quality of an electrophoretic transfer and act as a control in repetitive blotting experiments. The visibility of the standards makes it possible to monitor the separation of proteins while electrophoresis is in progress, even after the dye front has run off the gel. The standards can also be used to locate a protein for excision from an unstained prepar­ative gel.
Specifications
Contents Approximately 625 µg total protein*
in 33% (v/v) glycerol, 3% SDS, 10 mM Tris pH 7.0, 10 mM DTT, 2 mM EDTA, 0.01% NaN3.
Storage -20 °C Shelf life 1 year at -20 °C Volume 500 µl Applications 25-100
per vial
* Total protein concentration is an approximation based
on the average concentrations of multiple lots. Actual concentrations will vary.
Coomassie is a trademark of ICI.
21
Protein Molecular Weights
The molecular weights of every lot of prestained standards are individually calibrated against Bio-Rad’s SDS-PAGE Standards. The lot specific calibrated molecular weights are included with every vial. Prestained standards are useful for estimating the molecular weights of sample proteins, however, for precise molecu­lar weight determination, use Bio-Rad’s SDS­PAGE, Silver Stain SDS-PAGE, or Biotinylated SDS-PAGE Standards in addition to the prestained standards.
34
Instructions for Use
Heat the solution to 40 °C for 1 minute to dissolve any solids which may have pre­cipitated at -20 °C. To visualize the prestained standards after blotting, load 10 µ l for full size gels (16-20 cm) and 5 µl for mini gels. To visualize the standards during electrophoresis, load 20 µ l for full length gels and 10 µ l for mini gels. To see the stan­dards during the run, it is helpful to hold a sheet of white paper behind the gel.
Note: The prestaining of the proteins sub­stantially inhibits them from being further stained with biotin/avidin systems, colloidal gold, Coomassie standards can be silver stained, but silver staining will result in broad bands because of the large amount of protein in the sample.
®
R-250, or amido black. The
Constituent Proteins
(See enclosed insert for lot specific calibrated molecular weights.)
Protein Source
Phosphorylase B Rabbit muscle Bovine serum albumin Bovine plasma Ovalbumin Chicken egg white Carbonic anhydrase Bovine erythrocytes Soybean trypisn inhibitor Soybean Lysozyme Chicken egg white
† Covalently bound dye alters the molecular weight of the
proteins and produces relatively broad bands. The molecular weights are calibrated from the center of each protein band.
Fig. 1. Low Range Prestained SDS-PAGE Standards.
5 µl of the standards were run on a 12% SDS poly-acry­lamide gel according to the method of Laemmli. standards were run on the Mini-PROTEAN electrophoretically transferred to nitrocellulose using the Mini Trans-Blot
®
cell.
Phosphorylase B Bovine serum
albumin
Ovalbumin
Carbonic anhydrase
Soybean trypsin inhibitor
Lysozyme
®
II cell and
1
The
5 6
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