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    In vivo Leukemogenesis Model Using Retrovirus Transduction
    Authors:  Hiroshi Okuda and Akihiko Yokoyama, date: 12/05/2017, view: 80, Q&A: 0
    Various genetic alterations such as chromosomal translocation cause leukemia. For examples, gene rearrangements of the mixed-lineage leukemia (MLL) gene generate MLL fusion genes, whose products are potent oncogenic drivers in ...
    Myeloid Progenitor Transformation Assay
    Authors:  Hiroshi Okuda and Akihiko Yokoyama, date: 12/05/2017, view: 91, Q&A: 0
    Numerous oncogenes have been identified to cause leukemia. For example, chromosomal translocation generates various fusion genes of the mixed-lineage leukemia (MLL) gene and a partner gene in leukemia, whose gene products transform primary ...
    In vivo Leukemogenesis Model Using Retrovirus Transduction
    Authors:  Hiroshi Okuda and Akihiko Yokoyama, date: 12/05/2017, view: 80, Q&A: 0
    [Abstract] Various genetic alterations such as chromosomal translocation cause leukemia. For examples, gene rearrangements of the mixed-lineage leukemia (MLL) gene generate MLL fusion genes, whose products are potent oncogenic drivers in acute leukemia. To better understand the mechanism of disease onset, several murine leukemia models ...
    Myeloid Progenitor Transformation Assay
    Authors:  Hiroshi Okuda and Akihiko Yokoyama, date: 12/05/2017, view: 91, Q&A: 0
    [Abstract] Numerous oncogenes have been identified to cause leukemia. For example, chromosomal translocation generates various fusion genes of the mixed-lineage leukemia (MLL) gene and a partner gene in leukemia, whose gene products transform primary myeloid progenitors into an immortalized state. To characterize the transforming ability of leukemic ...
    An Affinity-directed Protein Missile (AdPROM) System for Targeted Destruction of Endogenous Proteins
    Authors:  Thomas J Macartney, Gopal P Sapkota and Luke J Fulcher, date: 11/20/2017, view: 311, Q&A: 0
    [Abstract] We recently reported an Affinity-directed PROtein Missile (AdPROM) system for the targeted proteolysis of endogenous proteins of interest (POI) (Fulcher et al., 2016 and 2017). AdPROM consists of the Von Hippel Lindau (VHL) protein, a Cullin 2 E3 ligase substrate receptor (Bosu and Kipreos, 2008), conjugated to a high affinity polypeptide ...
    In vitro NLK Kinase Assay
    Authors:  Sungho Moon, Jiyoung Kim and Eek‐hoon Jho, date: 11/05/2017, view: 351, Q&A: 0
    [Abstract] This protocol provides step by step instructions to perform an in vitro kinase assay for nemo-like kinase. In addition, this protocol also describes an efficient method using mild lysis buffer for expression and purification of Glutathione S-transferase (GST) fusion proteins.
    Xenograft Mouse Model of Human Uveal Melanoma
    Authors:  Yao Chen, Xiao Liu, Ling Gao and Yongqing Liu, date: 11/05/2017, view: 340, Q&A: 0
    [Abstract] Uveal melanoma (UM) is a malignant intraocular tumor in adults. Metastasis develops in almost half of the patients and over 90% of the metastases are in the liver. With the advances in molecular targeting therapy for melanoma, a proper metastasis animal model is of increasing importance for testing the accuracy and effectiveness of systemic ...
    Uptake Assays to Monitor Anthracyclines Entry into Mammalian Cells
    Authors:  Nicolas Brosseau, Emil Andreev and Dindial Ramotar, date: 09/20/2017, view: 580, Q&A: 0
    [Abstract] Anthracyclines, such as doxorubicin and daunorubicin, are DNA damaging agents that autofluoresce and can be readily detected in cells. Herein, we developed suitable assays to quantify and localize daunorubicin in mammalian cells. These assays can be exploited to identify components that are involved in the uptake of anthracyclines.
    Protocol for Establishing a Multiplex Image-based Autophagy RNAi Screen in Cell Cultures
    Authors:  Jennifer Jung and Christian Behrends, date: 09/05/2017, view: 821, Q&A: 0
    [Abstract] Autophagy is a recycling pathway, in which intracellular cargoes including protein aggregates and bacteria are engulfed by autophagosomes and subsequently degraded after fusion with lysosomes. Dysregulation of this process is involved in several human diseases such as cancer or neurodegeneration. Hence, advancing our understanding of how autophagy ...
    Isolation and Separation of Epithelial CD34+ Cancer Stem Cells from Tgfbr2-deficient Squamous Cell Carcinoma
    Authors:  Heather A. McCauley and Géraldine Guasch, date: 09/05/2017, view: 834, Q&A: 0
    [Abstract] Most epithelial tumors have been shown to contain cancer stem cells that are potentially the driving force in tumor progression and metastasis (Kreso and Dick, 2014; Nassar and Blanpain, 2016). To study these cells in depth, cell isolation strategies relying on cell surface markers or fluorescent reporters are essential, and the isolation ...
    GFP-Grb2 Translocation Assay Using High-content Imaging to Screen for Modulators of EGFR-signaling
    Authors:  Julia Petschnigg and Robin Ketteler, date: 09/05/2017, view: 717, Q&A: 0
    [Abstract] High-content screening is a useful tool to understand complex cellular processes and to identify genes, proteins or small molecule compounds that modulate such pathways. High-content assays monitor the function of a protein or pathway by visualizing a change in an image-based readout, such as a change in the localization of a reporter protein. ...
    [Bio101] Purifying Properly Folded Cysteine-rich, Zinc Finger Containing Recombinant Proteins for Structural Drug Targeting Studies: the CH1 Domain of p300 as a Case Example
    Authors:  Yong Joon Kim, Stefan Kaluz, Anil Mehta, Emily Weinert, Shannon Rivera and Erwin G. Van Meir, date: 09/05/2017, view: 824, Q&A: 0
    [Abstract] The transcription factor Hypoxia-Inducible Factor (HIF) complexes with the coactivator p300, activating the hypoxia response pathway and allowing tumors to grow. The CH1 and CAD domains of each respective protein form the interface between p300 and HIF. Small molecule compounds are in development that target and inhibit HIF/p300 complex formation, ...