• 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.4532

Current Issue in 2022

Volume: 12, Issue: 20

Biochemistry

Staphylococcus aureus 30S Ribosomal Subunit Purification and Its Biochemical and Cryo-EM Analysis Authors:  Margarita Belinite, Iskander Khusainov and Stefano Marzi, date: 10/20/2022, view: 118, Q&A: 0

The ribosome is a complex cellular machinery whose solved structure allowed for an incredible leap in structural biology research. Different ions bind to the ribosome, stabilizing inter-subunit interfaces and structurally linking rRNAs, proteins, and ligands. Besides cations such as K+ and Mg2+, polyamines are known to stabilize the folding of RNA

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Biological Engineering

A Novel Method of Inducible Directed Evolution to Evolve Complex Phenotypes Authors:  Ibrahim S. Al’Abri, Zidan Li, Daniel J. Haller and Nathan Crook, date: 10/20/2022, view: 174, Q&A: 0

Directed evolution is a powerful technique for identifying beneficial mutations in defined DNA sequences with the goal of improving desired phenotypes. Recent methodological advances have made the evolution of short DNA sequences quick and easy. However, the evolution of DNA sequences >5kb in length, notably gene clusters, is still a challenge

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Biophysics

Single Protein Detection and Imaging with Evanescent Scattering Microscopy Authors:  Pengfei Zhang, Lei Zhou, Rui Wang, Xinyu Zhou, Jiapei Jiang, Zijian Wan and Shaopeng Wang, date: 10/20/2022, view: 98, Q&A: 0

Single-molecule measurements provide statistical distributions of molecular properties, in addition to the ensemble averages. Evanescent detection approaches have been widely used for single-molecule detection because the evanescent field can significantly enhance the light-analyte interaction and reduce the background noise. However, current

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Single-molecule Force Spectroscopy on Biomembrane Force Probe to Characterize Force-dependent Bond Lifetimes of Receptor–ligand Interactions on Living Cells Authors:  Tongtong Zhang, Chenyi An, Wei Hu and Wei Chen, date: 10/20/2022, view: 146, Q&A: 0

The transmembrane receptor–ligand interactions play a vital role in the physiological and pathological processes of living cells, such as immune cell activation, neural synapse formation, or viral invasion into host cells. Mounting evidence suggests that these processes involve mechanosensing and mechanotransduction, which are directly

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Cancer Biology

Analysis of the Ubiquitination and Phosphorylation of Vangl Proteins Authors:  Di Feng, Ziwei He and Bo Gao, date: 10/20/2022, view: 95, Q&A: 0

The core planar cell polarity (PCP) protein Vang/Vangl, including Vangl1 and Vangl2 in vertebrates, is indispensable during development. Our previous studies showed that the activity of Vangl is tightly controlled by two important posttranslational modifications, ubiquitination and phosphorylation. Vangl is ubiquitinated through an endoplasmic

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Cell Biology

Measuring Intracellular H2O2 in Intact Human Cells Using the Genetically Encoded Fluorescent Sensor HyPer7 Authors:  Lianne J. H. C. Jacobs, Michaela N. Hoehne and Jan Riemer, date: 10/20/2022, view: 69, Q&A: 0

Depending on its local concentration, hydrogen peroxide (H2O2) can serve as a cellular signaling molecule but can also cause damage to biomolecules. The levels of H2O2 are influenced by the activity of its generator sites, local antioxidative systems, and the metabolic state of the cell. To study and understand the role of H2O2 in cellular

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Developmental Biology

First-line Evaluation of Sperm Parameters in Mice (Mus musculus) Author:  Guillaume Martinez, date: 10/20/2022, view: 74, Q&A: 0

Infertility has become a major public health problem, with a male factor involved in about half the cases. Mice are the most widely used animal model in reproductive biology research laboratories, but changes in sperm parameters in mice can be subtle and, in the absence of official guidelines, it is important that analyses are carried out in a

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Microbiology

Negative Staining Transmission Electron Microscopy of HIV Viral Particles Permeabilized with PFO and Capsid Stabilized with IP6 Authors:  Derrick Lau, Chantal L. Márquez, Michael W. Parker and Till Böcking, date: 10/20/2022, view: 76, Q&A: 0

The human immunodeficiency virus 1 (HIV-1) consists of a viral membrane surrounding the conical capsid. The capsid is a protein container assembled from approximately 1,500 copies of the viral capsid protein (CA), functioning as a reaction and transport chamber for the viral genome after cell entry. Transmission electron microscopy (TEM) is a

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Neuroscience

MiniSOG2-mediated Specific Photoablation of Motor Neurons in Ascidian Embryos Authors:  Madoka K. Utsumi, Taichi Akahoshi, Kotaro Oka and Kohji Hotta, date: 10/20/2022, view: 128, Q&A: 0

When understanding the neuronal function of a specific neural circuit, single-cell level photoablation of a targeted cell is one of the useful experimental approaches. This protocol describes a method to photoablate specific motor neurons via the mini singlet oxygen generator (miniSOG2), a light–oxygen–voltage (LOV)-based optogenetic

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Plant Science

Measurement of Ascorbate Peroxidase Activity in Sorghum Author:  Praveen Kumar, date: 10/20/2022, view: 67, Q&A: 0

The ascorbate peroxidase (APX) is a widely distributed antioxidant enzyme. It differs from catalase and other peroxidases in that it scavenges/reduces reactive oxygen species (ROS) such as hydrogen peroxide (H2O2) to water using reduced ascorbate as the electron donor. It is advantageous over other similar antioxidant enzymes in scavenging ROS

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