By P. Michael Conn
This new quantity of Methods in Enzymology keeps the legacy of this prime serial with caliber chapters authored by means of leaders within the box. this is often the second of 2 volumes on endosome signaling and contains chapters on such issues as size of access into the endosomal compartment via multi-parametric photograph research, review of peptide internalization and endosomal signaling, and VEGF-A in endosomal signaling.
- Continues the legacy of this leading serial with caliber chapters authored through leaders within the box
- Covers endosome signaling
- Contains chapters on such themes as dimension of organic results of endosomal proteolysis of internalized insulin and multi-vesicular endosome biogenesis.
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Continue this process until all four plates per given condition have been lysed. 5 mL) to a 15 mL conical tube. The cell lysate should be very viscous. 6. Repeat the previously mentioned lysis procedure for each of the other conditions and place into separate 15 mL conical tubes. 7. Homogenize the cell lysate with three 10-s bursts in a Polytron tissue grinder (settings 7–8). It is recommended that the samples be kept on ice between homogenizations. Following homogenization, sonicate samples with three 20-s bursts of sonication on ice.
In addition, several mutations have been introduced into the original YFP to accelerate maturation, which leads to the production of brighter YFP proteins (Kodama & Hu, 2012). , 2002), which contains a F46L mutation in the backbone of the “super”-EYFP (SEYFP) (Sawano & Miyawaki, 2000). The Venus is not only brighter than SEYFP but also more resistant to acid and ClÀ, which are common in many cellular compartments. In our study (Chen, Liu, & Songyang, 2007), the Yn is an N-terminal fragment of Venus, truncated at residue 155, a common split site for BiFC (Kodama & Hu, 2012).
Molecular Oncology, 3, 297–307. , & Yamamoto, M. (1988). Isolation and characterization of Schizosaccharomyces pombe mutants phenotypically similar to ras1. Molecular & General Genetics, 215, 26–31. , et al. (1997). Protein binding and signaling properties of RIN1 suggest a unique effector function. In: Proceedings of the National Academy of Sciences of the United States of America, 94, 4954–4959. Hu, C. , & Kerppola, T. K. (2002). Visualization of interactions among bZIP and Rel family proteins in living cells using bimolecular fluorescence complementation.
Endosome Signaling Part B by P. Michael Conn