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    • What Is Epigenetics?

      If all cells are created from the same genetic material, why are there so many different cell types? Listen to Sriharsa Pradhan, Senior Scientist, RNA Biology at NEB, as he describes how DNA is methylated and how this affects the path of reading the DNA code the same way an obstruction would derail a train off its tracks.

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    • A Breakthrough Method of RNA Sample Preparation

      The common problem of adaptor dimer formation during small RNA library construction can be avoided by using NEBNext® protocols. Learn about this technique, and how it improves both performance and sensitivity in library construction.

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    • SNAP-tag® Overview Tutorial

      View an interactive tutorial explaining the mechanism of our SNAP-tag® technologies and reagents available for researchers wishing to study the function and localization of proteins in live or fixed cells.

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    • Overview of Glycobiology

      Learn about the core sequences and common modifications of N-linked and O-linked glycans in this video. Analysis of these glycans and/or peptide portions of the glycoprotein can be accomplished with the use of deglycosylation enzymes, which are explained in detail. Unlike other chemical deglycosylation methods, enzymatic treatment is much gentler and can provide complete sugar removal with no protein degradation.

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    Epigenetics

    Epigenetics is the study of heritable changes in gene expression that are not encoded in the DNA of the genome. Encouraging evidence has linked epigenetic effects to oncogenesis, progression and treatment of cancer (1), the regulation of development and function of the nervous system (2), gene regulation (3), cellular stress events (3), nutrigenomics (4), aging and DNA repair (5). Considerable ongoing efforts are directed towards identifying the dynamic functions of various modifications to DNA and its associated proteins and elucidating their mechanisms.

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    References

    1. Baylin, S. and Jones, P. (2011) Nature Rev .Cancer 11(10): 726-734. PMID: 21941284
    2. Riccio, A., (2010) Nature Neuroscience, 13: 1330-1337. PMID: 20975757
    3. Huang, J. et al. (2006) Nature, 444: 629-632. PMID: 17108971
    4. Park, L.K., Friso, S. and Choi, S.W. (2011) Proc. Nutr. Soc. 4: 1-9.
    5. Pahlich, S., Zakaryan, R.P. and Gehring., H. (2006) Biochim. Biophys. Acta. 1764: 1890-1903. PMID: 17010682 

    Glycobiology

    Glycobiology is the study of the structure, function and biology of carbohydrates, often called glycans, which are widely distributed in nature (1). It is a small but rapidly growing field in biology with relevance to biomedicine, biotechnology, biofuels and basic research. In eukaryotic cells, the majority of proteins are post-translationally modified (2). A common modification, essential for cell viability, is the attachment of glycans. 

    Glycans define many properties of glycoconjugates (glycoproteins and glycolipids). For instance, it is largely through glycan–protein interactions that cell–cell and cell–pathogen contacts occur. Likewise, glycan molecules modulate many other processes important for cell and tissue differentiation, metabolic and gene regulation, protein activity, protein clearance, transport and more (3-10). 


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    References

    1. Spiro, R.G. (2002) Glycobiology 12: 43R-56R. PMID: 112042244 
    2. Khoury, G.A. et al. (2011) Scientific Reports 1: 90. PMID: 22034591  
    3. Varki, A. (1993) Glycobiology, 3(2): 97-130. PMID: 8490246 
    4. Zhao, Y.Y. et al.  (2008) Cancer Sci. 99(7): 1304-1310. PMID: 18492092 
    5. Zhao, Y. et al.  (2008) FEBS J. 275(9): 1939-1948. PMID: 18384383  
    6. Skropeta, D. (2009) Bioorg Med Chem. 17(7): 2645-2653. PMID: 19285412 
    7. Neu, U. et al.  (2011) Curr Opin Struct Biol. 21(5): 610-618. PMID: 21917445 
    8. Cerliani, J.P. et al.  (2011) J Clin Immunol. 31(1): 10-21. PMID: 21184154 
    9. Aarnoudse, C.A. et al.  (2006) Curr Opin Immunol. 18(1): 105-111. PMID: 16303292 
    10. Arnold, J.N. (2006) Immunol Lett. 106(2): 103-110. PMID: 1681439 

    Protein Expression and Purification

    Recombinant production of proteins is one of the most powerful techniques used in the Life Sciences. The ability to produce and purify an abundance of a desired recombinant protein can permit a wide range of possibilities including its biochemical or structural characterization, its use in industrial processes or its use to diagnose or treat disease.

    At first glance, recombinant protein expression looks quite simple. Essentially, DNA encoding a target protein is cloned downstream of a promoter in an expression vector. This vector is then introduced into a host cell, and the cell’s protein synthesis machinery produces the desired protein. In practice, however, protein expression can be very challenging because so many factors may influence the process. For example, each protein folds in its own unique manner, a process that may be influenced by the choice of expression host. Similarly, some proteins require post-translational modifications or proper insertion into a biological membrane. Finally, some proteins may have an activity that is detrimental to the host. Thus, no single solution exists for successful production of all recombinant proteins. Instead, it is beneficial to have access to a wide range of expression tools and a willingness to explore multiple approaches to better one’s chances for success.

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    Protein Analysis and Tools

    Not only are proteins a major structural and catalytic component of living systems, they can also be effector molecules whose states determine downstream activities. Therefore, studying the protein complement within a cell can reveal the mechanisms behind many of the cell’s responses to its environment. Given the vast number of applications for protein analysis, several tools and methods for its study exist; determining the correct method for your application is paramount to success.


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    RNA Analysis

    RNA is a polynucleotide that is typically transcribed from a DNA template. Historically, its only known function was involvement in the translation of genetic information into proteins, either through encoding the sequence of the protein (messenger RNA; mRNA), bringing amino acids to the ribosome (transfer RNA; tRNA) or by being a component of the ribosome translational machinery (ribosomal RNA; rRNA). Recently, technological advances and innovative research in the field of RNA analysis have led to the discovery of many new classes of small and large non-coding RNAs with novel regulatory functions. These findings have helped usher in a renaissance of RNA-focused research in biology.

    Key capabilities for contemporary RNA research include RNA synthesis for in vitro analysis or transfection, RNA modification and processing, reverse transcription of RNA for cloning and expression analysis and detection of RNA.

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    Cellular Analysis

    Studies on protein properties, expression, transport, degradation and their interactions with other cellular systems are integral to biomedical research, drug discovery and developmental biology. Eukaryotic proteins function in signaling pathways, metabolism, structure, adhesion, cell movement, active and passive transport, DNA repair, viral disease mechanisms, the immune system, fertilization, differentiation, epigenetics, cancer and the cell division cycle.

    For efficient analysis of cellular protein interactions and expression, target genes are often engineered into reporter systems, then expressed in cells as recombinant proteins. Reporter gene characteristics are chosen to enable downstream in vitro or in vivo method applications. This commonly used and widely successful strategy can facilitate by applications such as flow cytometry, cell sorting, in vitro or in vivo imaging, proteomic microarray, cell-based microarray, SDS-PAGE gel electrophoresis, western blot, quantitative affinity purification followed by mass spectrometry (q-AP-MS), pulse-chase and receptor internalization assays. In particular, improved reporter systems in conjunction with optical imaging technical advances are pushing the frontiers of fluorescence imaging applications.

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