HI
Hogune Im
  • Molecular and Cellular Pharmacology Program, Department of Pharmacology, University of Wisconsin Medical Schoo, USA
Research fields
  • Biochemistry
Restriction Enzyme Accessibility Protocol in Mammalian Cells
Author:  Hogune Im, date: 10/20/2011, view: 11918, Q&A: 0
This protocol describes a method to indirectly assess chromatin structure by using restriction enzyme in mammalian cells, modified from Current Protocols.
Mouse Embryonic Stem Cell Maintenance for Differentiation
Author:  Hogune Im, date: 10/20/2011, view: 16964, Q&A: 0
Embryonic stem cells are derived from inner cell mass of an embryo that can differentiate into every cell type in the body. Clinically, cultured red blood cell supply is of great interest. However, some of the hurdles need to be overcome. This protocol describes a protocol to maintain mouse stem cells that will be used for hematopoietic lineage differentiation assay in the lab.
Mouse Embryonic Stem Cell Differentiation to Hematopoietic Precursors
Author:  Hogune Im, date: 04/05/2012, view: 15188, Q&A: 0
Embryonic stem cells are derived from inner cell mass of an embryo that can differentiate into every cell type in the body. Clinically, cultured red blood cell supply is of great interest. However, some of the hurdles need to be overcome. This protocol describes a two step protocol to form embryoid body and differentiate them in to hematopoietic lineage cells, especially erythroid cells.
The Inoue Method for Preparation and Transformation of Competent E. coli: "Ultra Competent" Cells
Author:  Hogune Im, date: 10/20/2011, view: 47556, Q&A: 1
This protocol differs from other procedures in that the bacterial culture is grown at 18 °C rather than the conventional 37 °C. Otherwise, the protocol is unremarkable and follows a fairly standard course. Why growing the cells at low temperature should affect the efficiency of transformation is unknown. Perhaps the composition or the physical characteristics of bacterial membranes synthesized at 18 °C are more favorable for uptake of DNA, or perhaps the phases of the growth cycle that favor efficient transformation are extended. Incubating bacterial cultures at 18 °C is a challenge. Most laboratories do not have a shaking incubator that can accurately maintain a temperature of 18 °C summer and winter. One solution is to place an incubator in a 4 °C cold room and use the temperature control to heat the incubator to 18 °C. Alternatively, there is almost no loss of efficiency if the cultures are grown at 20-23 °C, which is the ambient temperature in many laboratories. Cultures incubated at these temperatures grow slowly with a doubling time of 2.5 to 4 h. To avoid reaching desired OD late at night, set up cultures in the evening and harvest the bacteria early the following morning. The procedure works well with many strains of E. coli in common use in molecular cloning, including XL1-Blue, DH1, JM103, JM108/9, DH5a, and HB101.
GST-tagged Yeast Protein Purification
Author:  Hogune Im, date: 10/05/2011, view: 16881, Q&A: 2
Glutation S-transferase (GST) tagging is the most commonly used purification strategy for recombinant protein. It was developed with the goal of preserving the enzymatic activity by utilizing gentle elution condition of the target protein from purification matrix (Poon and Hunt., 1994). The method described here can be applied from single protein to proteome scale purification of recombinant protein from yeast (Zhu et al., 2000; Zhu et al., 2001).
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