• Volume 12, 2022
  • Volume 11, 2021
  • Volume 10, 2020
  • Volume 9, 2019
  • Volume 8, 2018
  • Volume 7, 2017
  • Volume 6, 2016
  • Volume 5, 2015
  • Volume 4, 2014
  • Volume 3, 2013
  • Volume 2, 2012
  • Volume 1, 2011
Image Credit: BioProtoc.4500

Past Issues in 2022

Volume: 12 Issue: 17

Biochemistry

Split-Chloramphenicol Acetyl Transferase Assay to Study Protein-Protein Interactions and Ubiquitylation in Escherichia coli Authors:  Amir Florentin, Alina Kordonsky, Elon Yariv, Reut Avishid, Noa Efron, Edache Akogwu and Gali Prag, date: 09/05/2022, view: 429, Q&A: 0

Protein-protein interactions and protein modifications play central roles in all living organisms. Of the more than 200 types of post-translational modifications, ubiquitylation is the most abundant, and it profoundly regulates the functionality of the eukaryotic proteome. Various in vitro and in vivo methodologies to study protein interactions

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Production, Purification, and Fluorometric Activity Assay of Human Aldehyde Dehydrogenases Authors:  Raquel Pequerul, Sergio Porté, Xavier Parés, Mileidys Pérez-Alea and Jaume Farrés, date: 09/05/2022, view: 385, Q&A: 0

Human aldehyde dehydrogenase (ALDH) isoforms are NAD(P)+-dependent enzymes catalyzing the oxidation of a wide spectrum of aldehydes to their corresponding carboxylic acids. ALDH families 1 and 3 are associated with stemness, chemoresistance, and tumor progression across multiple tumor types. ALDH2 is involved in ethanol metabolism, and its

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Biological Engineering

Incorporation of a Chemically Diverse Set of Non-Standard Amino Acids into a Gram-Positive Organism Authors:  Devon A. Stork, Michaela A. Jones, Ethan C. Garner and Aditya M. Kunjapur, date: 09/05/2022, view: 317, Q&A: 0

The incorporation of non-standard amino acids (nsAAs) within proteins and peptides through genetic code expansion introduces novel chemical functionalities such as photo-crosslinking and bioconjugation. Given the utility of Bacillus subtilis in fundamental and applied science, we extended existing nsAA incorporation technology from Escherichia

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Cancer Biology

Microscopic Detection of DNA Synthesis in Early Mitosis at Repetitive lacO Sequences in Human Cells Authors:  Kazumasa Yoshida, Riko Ishimoto and Masatoshi Fujita, date: 09/05/2022, view: 571, Q&A: 0

In the human cell cycle, complete replication of DNA is a fundamental process for the maintenance of genome integrity. Replication stress interfering with the progression of replication forks causes difficult-to-replicate regions to remain under-replicated until the onset of mitosis. In early mitosis, a homology-directed repair DNA synthesis,

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Detection of Alternative End-Joining in HNSC Cell Lines Using DNA Double-Strand Break Reporter Assays Authors:  Nan Zuo, Lin Ma, Weitao Hu, Yongqiang Deng, Lanlan Wei and Qi Liu, date: 09/05/2022, view: 640, Q&A: 0

The main cellular pathways to repair DNA double-strand breaks (DSBs) and protect the integrity of the genome are homologous recombination (HR), non-homologous end-joining (NHEJ), and alternative end-joining (Alt-EJ). Polymerase theta-regulated Alt-EJ is an error-prone DSB repair pathway characterized by microhomology usage. Considering its

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Immunology

High-Efficiency Retroviral Transduction for Type 1 Regulatory T Cell Differentiation Authors:  Michael C. McGee, Avery August and Weishan Huang, date: 09/05/2022, view: 365, Q&A: 0

Type 1 regulatory T (Tr1) cells are an immunoregulatory CD4+ Foxp3- IL-10high T cell subset with therapeutic potential for various inflammatory diseases. Retroviral (RV) transduction has been a valuable tool in defining the signaling pathways and transcription factors that regulate Tr1 differentiation and suppressive function. This protocol

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Microbiology

A New Tool for the Flexible Genetic Manipulation of Geobacillus kaustophilus Authors:  Ryotaro Amatsu, Kotaro Mori, Shu Ishikawa, Wilfried J. J. Meijer and Ken-ichi Yoshida, date: 09/05/2022, view: 308, Q&A: 0

Geobacillus kaustophilus, a thermophilic Gram-positive bacterium, is an attractive host for the development of high-temperature bioprocesses. However, its reluctance against genetic manipulation by standard methodologies hampers its exploitation. Here, we describe a simple methodology in which an artificial DNA segment on the chromosome of

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Molecular Biology

Carbamoyltransferase Enzyme Assay: In vitro Modification of 5-hydroxymethylcytosine (5hmC) to 5-carbamoyloxymethylcytosine (5cmC) Authors:  Weiwei Yang, Nan Dai, Yu-Cheng Lin, William Johnson, Romualdas Vaisvila, Peter Weigele, Yan-Jiun Lee, Ira Schildkraut, Ivan R. Corrêa Jr. and Laurence Ettwiller, date: 09/05/2022, view: 304, Q&A: 0

Nucleic acids in living organisms are more complex than the simple combinations of the four canonical nucleotides. Recent advances in biomedical research have led to the discovery of numerous naturally occurring nucleotide modifications and enzymes responsible for the synthesis of such modifications. In turn, these enzymes can be leveraged towards

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Neuroscience

Evaluation of Mitochondrial Turnover Using Fluorescence Microscopy in Drosophila Author:  Felipe Martelli, date: 09/05/2022, view: 408, Q&A: 0

Mitochondrial dysfunction is associated with perturbations in the cellular oxidative status, changes in energy production and metabolic rate, and the onset of pathological processes. Classic methods of assessing mitochondrial dysfunction rely on indirect measures, such as evaluating mitochondrial DNA copy numbers, or direct but more costly and

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Stem Cell

Generation of iMyoblasts from Human Induced Pluripotent Stem Cells Authors:  Dongsheng Guo, Katelyn Daman, Danielle Fernandes Durso, Jing Yan and Charles P. Emerson Jr., date: 09/05/2022, view: 418, Q&A: 0

Skeletal muscle stem cells differentiated from human-induced pluripotent stem cells (hiPSCs) serve as a uniquely promising model system for investigating human myogenesis and disease pathogenesis, and for the development of gene editing and regenerative stem cell therapies. Here, we present an effective and reproducible transgene-free protocol for

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