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In JoVE (1)
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Articles by Mark Laible in JoVE
JoVE General
Домашний сайт направленного мутагенеза Всего Плазмиды
1Department of Biology, Johannes Gutenberg-University Mainz, Germany, 2Proteomics division, AlPlanta, Neustadt an der Weinstrasse, Germany
Сайт направленного мутагенеза целых плазмид простой способ создать несколько различных вариантов оригинальных плазмиды. Здесь мы показываем, простой и экономически эффективный способ представить база замен в плазмиду с использованием реагентов стандарта.
Other articles by Mark Laible on PubMed
Contact Spotting of Protein Microarrays Coupled with Spike-in of Normalizer Protein Permits Time-resolved Analysis of ERBB Receptor Signaling
Protein microarrays allow highly accurate comparison and quantification of numerous biological samples in parallel while requiring only little material. This qualifies protein arrays for systems biology and clinical research where only limited sample material is available, but a precise readout is required. With the introduction of signal normalization steps to monitor the drop size of manually contact-spotted RP protein arrays, the usefulness of normalizer proteins to ensure a high-throughput but inexpensive protein analysis was demonstrated. This approach was applied for the analysis of signaling through ERBB receptor activated kinases in the breast cancer cell line MCF-7. Activation of ERK1/2 and AKT by ERBB1 (EGFR), ERRB2 (HER2/neu), and ERBB3-4 was monitored in a time-resolved manner. Analysis of pathway activation by stimulation with epidermal growth factor and heregulin, or inhibition by blocking with gefitinib or herceptin allowed a characterization of the distinct signaling properties of the different ERBB receptor subtypes.
