![]() All the ladder proteins can be stained using dyes such as Coomassie Blue and are also directly detectable in Western blots through the incorporation of immunoglobulin antibody binding domains. This customization is easy to accomplish with the Penn State Protein Ladder system”įor even more efficient production, the ladders are also available as two coexpression vectors that produce either the set of 10, 30, 50, 100 kD proteins or the set of 20, 40, 60, 80 kD proteins. “They might not need the entire set, or they might want to individually control the intensity of bands produced on a gel. “Researchers can pick and choose amongst the nine individual proteins to design a ladder optimized for their research project,” said Tan. 50 milliliters (ml) of cells produce enough of each individual protein for 20,000 experiments. coli cells containing the plasmids and then purify the proteins using a common affinity tag designed into each protein. Using standard laboratory protocols, researchers can grow E. Proteins can be detected on Western blots by staining with Ponceau S, Coomassie Blue, (except for 3.4kDa and 5kDa polypeptides) or by using Strep- Tactin conjugates. Each protein is encoded on an individual plasmid-a circular form of DNA-that is expressed at very high levels in the bacteria E. SDS-PAGE 5: Interpreting Results from a Protein Gel (periplasmic extract from E. These standards contain ten Strep-tagged recombinant proteins (10250 kD), including three pink reference bands (25, 50, and 75 kD), as well as unstained versions of the Strep-tagged recombinant proteins for molecular weight sizing after blot development. Precision Plus Protein Dual Color Standards 500 L 1610374. The Penn State Protein Ladder is composed of nine proteins that range in molecular weight from 10 to 100 kilodaltons (kD). Use Precision Plus Protein WesternC Standards as a combination standard for western blotting and fluorescence detection in SDS-PAGE. “Our Penn State Protein Ladders can be easily made for a tiny fraction of that cost using source material that we make available to nonprofit academic researchers through the Addgene and DNASU plasmid repositories.” Review other protein ladders in the unstained and prestained protein ladder guide. Recommended loading: 2 - 3 µl for western blots. Easy to identify: Includes green 25 kDa and red 75kDa reference bands. Ready-to-use: Supplied in a loading buffer for direct loading on gels. “Any lab working with proteins uses these ladders on a daily basis and the cost for commercially available ladders adds up-averaging about $1.00 per experiment,” said Tan. Key product features: Broad range: 10-180 kDa. Protein ladders are molecular rulers for estimating the sizes of proteins separated by gel electrophoresis. “It has been exciting and rewarding to develop this project from just an idea to tools that can serve science,” said Williamson. We recommend using unstained protein ladders for molecular weight estimation applications as prestained ladders have a dye that is covalently bound to each protein that will result in the ladder migrating differently in different buffer systems (i.e., different gels). Fleischman, all of whom were Penn State undergraduate students. Williamson III, Yoshitaka Shibata, Rosalie P. Willaman Professor of Molecular Biology at Penn State, developed the ladders to be easily used in two of the most common experiments-SDS-PAGE and Western blots-in protein research.Ī paper describing the research appears Augin the journal Scientific Reports. ![]() Improved Dual Color Standards are brighter than the competition on both gels and blots.Labs can easily make their own protein ladders for less than a penny per experiment using the newly developed, license-free “Penn State Protein Ladder system.” A research team of undergraduate students led by Song Tan, Verne M. Increased overall brightness provides visual robustness throughout the entire protein electrophoresis and western blotting workflow.
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