Volume: 5 Issue: 22 —BIO-PROTOCOL

Cancer Biology

¹³C Tracer Studies of Metabolism in Mouse Tumor Xenografts Authors: Andrew N. Lane, Jun Yan and Teresa W-M. Fan, date: 11/20/2015, view: 8294, Q&A: 0

Mice are widely used for human tumor xenograft studies of cancer development and drug efficacy and toxicity. Stable isotope tracing coupled with metabolomic analysis is an emerging approach for assaying metabolic network activity. In mouse models there are several routes of tracer introduction, which have particular advantages and disadvantages ...

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Telomere Dysfunction Induced Foci (TIF) Analysis Authors: Ilgen Mender and Jerry W. Shay, date: 11/20/2015, view: 7789, Q&A: 0

Telomerase maintains telomeric DNA in eukaryotes during early developments, ~90% of cancer cells and some proliferative stem like cells. Telomeric repeats at the end of chromosomes are associated with the shelterin complex. This complex consists of TRF1, TRF2, Rap1, TIN2, TPP1, POT1 which protect DNA from being recognized as DNA double-stranded ...

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Telomerase Repeated Amplification Protocol (TRAP) Authors: Ilgen Mender and Jerry W. Shay, date: 11/20/2015, view: 13860, Q&A: 1

Telomeres are found at the end of eukaryotic linear chromosomes, and proteins that bind to telomeres protect DNA from being recognized as double-strand breaks thus preventing end-to-end fusions (Griffith et al., 1999). However, due to the end replication problem and other factors such as oxidative damage, the limited life span of cultured ...

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Telomere Restriction Fragment (TRF) Analysis Authors: Ilgen Mender and Jerry W. Shay, date: 11/20/2015, view: 15016, Q&A: 0

While telomerase is expressed in ~90% of primary human tumors, most somatic tissue cells except transiently proliferating stem-like cells do not have detectable telomerase activity (Shay and Wright, 1996; Shay and Wright, 2001). Telomeres progressively shorten with each cell division in normal cells, including proliferating stem-like cells, due to ...

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In vitro and in vivo Limiting Dilution Assay for Colorectal Cancer Authors: Lauren Agro and Catherine A O’Brien, date: 11/20/2015, view: 18038, Q&A: 0

The in vitro limiting dilution assay is used to determine the colorectal cancer initiating cell (CC-IC) frequency of a CC-IC enriched suspension culture, grown in growth factor enriched serum free media. The in vivo limiting dilution assay is used to determine the colorectal cancer initiating cell frequency of a primary ...

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Immunology

Skin Wound Healing Model - Excisional Wounding and Assessment of Lesion Area Authors: Camila Francisco Moreira, Puebla Cassini-Vieira, Mariana Felipetto da Silva and Lucíola da Silva Barcelos, date: 11/20/2015, view: 16703, Q&A: 5

This protocol focus on the most common surgical mouse model of cutaneous excisional wound healing used to study the cellular and molecular pathways involved in wound repair and regeneration as well as in translational applications such as the evaluation of new therapeutic modalities. This model allows the monitoring of the wound closure and the ...

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Estimation of Wound Tissue Neutrophil and Macrophage Accumulation by Measuring Myeloperoxidase (MPO) and N-Acetyl-β-D-glucosaminidase (NAG) Activities Authors: Puebla Cassini-Vieira, Camila Francisco Moreira, Mariana Felipetto da Silva and Lucíola da Silva Barcelos, date: 11/20/2015, view: 9975, Q&A: 0

The inflammatory response is essential to the reestablishment of cutaneous homeostasis following injury. In this context, leukocytes arrive at the wound site and orchestrate essential events in the wound healing process. Therefore, the quantification of specific subsets of inflammatory cells in the wound tissue is of considerable interest. The ...

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Permanent Occlusion of the Left Anterior Coronary Artery in the Rat Authors: Mélanie Bousquenaud, Pierre-Yves Marie and Yvan Devaux, date: 11/20/2015, view: 6143, Q&A: 0

Left ventricular (LV) remodeling occurs in many patients after myocardial infarction (MI). LV remodeling is characterized by progressive ventricular dilatation and contractile dysfunction, consequently to cardiomyocyte hypertrophy and fibrosis. Despite reperfusion therapies, this pathophysiological process is the main cause of cardiac evolution ...

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Microbiology

An Assay to Test the Capacity of Arabidopsis Plant Defensin Type1 Protein to Induce Cellular Zinc (Zn) Tolerance in Yeast Authors: Zaigham Shahzad, Laurence Marquès, Pierre Berthomieu and Françoise Gosti, date: 11/20/2015, view: 5708, Q&A: 0

Heterologous expression of genes in budding yeast Saccharomyces cerevisiae (S. cerevisiae) is especially suitable to functionally study the corresponding encoded protein at the cellular level (Bonneaud et al., 1991). This is mainly because many strains defective in specific activities are available and could be ...

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Purification of Bacterial RNA from Infected Macrophages Authors: Lior Lobel, Nadejda Sigal, Anna Pasechnek and Anat A. Herskovits, date: 11/20/2015, view: 7588, Q&A: 0

Studying the transcriptome of bacterial pathogens during infection is a very informative and effective tool for discovering genes that contribute to successful infection. However, isolating bacterial RNA from infected cells or tissues is a challenging process due to the much higher amounts of host RNA in the lysates of infected cells. We have ...

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Neuroscience

Locomotor Coordination Assay in Rats Authors: J. Regino Perez-Polo, Harriet C. Rea and Smitha K. Infante, date: 11/20/2015, view: 6250, Q&A: 0

Neonatal hypoxia-ischemia (HI) affects 60% of low birth weight infants and up to 40% of preterm births. Cell death and brain injury after HI have been shown to cause long-lasting neurological deficits. Two motor coordination tests on rats that had been exposed to HI on postnatal day 7 (P7) showed that HI in the P7 rat is associated with ...

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Plant Science

Expression, Purification and in vitro Enzyme Activity Assay of Plant Derived GTPase Authors: Annemarie Glöckner and Christian A. Voigt, date: 11/20/2015, view: 6334, Q&A: 0

Based on gene expression data after biotic stress, the GTPase RabA4c has been suggested to regulate pathogen-induced callose biosynthesis in the model organism Arabidopsis thaliana. We studied the function of RabA4c in its native and dominant negative (dn) isoform. In planta, RabA4c overexpression ...

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Quantifying the Permeability of the Apoplastic Water Barrier in Cosmos Petals Authors: Reinhard Jetter and Christopher Buschhaus, date: 11/20/2015, view: 5028, Q&A: 0

The capacity of plants to minimize uncontrolled water loss is essential for survival in adverse and changing climatic conditions. In order to assess and compare the effectiveness of apoplastic barriers to water, the permeability of the barrier must first be quantified. Studies have accomplished this directly by quantifying tritium flux or ...

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Arabidopsis Leaf Explant Culture Authors: Jing-Jing Wang, Lisha Zhang and Hui-Shan Guo, date: 11/20/2015, view: 8012, Q&A: 0

In this protocol, Arabidopsis leaf explant culture is described using an adaptation of a previous method (Hu et al., 2000). Cells from the cut edges of leaf explant are able to proliferate and subsequently form calli on the callus induction medium, in which is supplemented with 2,4-D and 6-benzyl aminopurine [6-BA]. 2,4-D, one of ...

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Sample Preparation and Fractionation of Arabidopsis thaliana Sperm and Vegetative Cell Nuclei by FACS Authors: Nina Chumak, Magdalena Mosiolek and Vera Karolina Schoft, date: 11/20/2015, view: 8993, Q&A: 1

One of the major topics in plant and animal biology is sexual reproduction. It is, therefore, of great interest to isolate and study germ cells and accessory cells. The male gametophyte of the flowering plant Arabidopsis thaliana (A. thaliana), pollen, is the product of two post-meiotic mitotic divisions. Each mature pollen grain ...

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Past Issues in 2015

Volume: 5 Issue: 22

Cancer Biology

Immunology

Microbiology

Neuroscience

Plant Science