• 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.4192

Biochemistry

Co-immunofluorescence of MRPL12 and Nrf2 in HK2 Cells Authors:  Yao Lu, Xia Gu, Qiang Wan, Hong Feng and Yi Liu, date: 10/20/2021, view: 446, Q&A: 0

Immunofluorescence is a technique to visualize the localization of specific molecule targets within cells using the specificity of antibodies. Here, we describe a protocol to detect two different protein components in a cell simultaneously. Antibody concentrations to be used vary from cell to cell and should be optimized for different cell types.

...
More >>

Biophysics

Measurement of the Translational Diffusion Coefficient and Hydrodynamic Radius of Proteins by Dynamic Light Scattering Authors:  Ernest T. Parker and Pete Lollar, date: 10/20/2021, view: 725, Q&A: 0

Diffusion is a fundamental process in biological systems that governs the molecular collisions driving biochemical reactions and membrane and transport. Measurement of the diffusion coefficient and application of the Stokes-Einstein equation produces the hydrodynamic radius, which is a commonly used gauge of particle size. Additionally,

...
More >>
Electrophysiological Recordings of the Polycystin Complex in the Primary Cilium of Cultured Mouse IMCD-3 Cell Line Authors:  Kotdaji Ha and Markus Delling, date: 10/20/2021, view: 371, Q&A: 0

PC-1 and PC-2 form an ion channel complex called the polycystin complex, which predominantly localizes to a small hair-like organelle called the primary cilium. The polycystin complex permeates cations, K+, Na+, and Ca2+, and has an unusual 1:3 stoichiometry that combines one PC-1 subunit with three PC-2 subunits. However, the small size and shape

...
More >>
Biophysical Characterization of Iron-Sulfur Proteins Authors:  Bhanu P. Jagilinki, Irina Paluy, Alexei M. Tyryshkin, Vikas Nanda and Dror Noy, date: 10/20/2021, view: 408, Q&A: 0

Iron-sulfur proteins are primordial catalysts and biological electron carriers that today drive major metabolic pathways across all forms of life. They can access a diversity of oxidation states and can mediate electron transfer over an extended range of reduction potentials spanning more than 1 V. Depending on the protein micro-environment and

...
More >>

Cancer Biology

Urea Denaturation, Zinc Binding, and DNA Binding Assays of Mutant p53 DNA-binding Domains and Full-length Proteins Authors:  Jeung-Hoi Ha, Xin Yu, Darren R. Carpizo and Stewart N. Loh, date: 10/20/2021, view: 365, Q&A: 0

In the cell, the thermodynamic stability of a protein – and hence its biological activity – can change dramatically as a result of perturbations in its amino acid sequence and the concentration of stabilizing ligands. This interplay is particularly evident in zinc-binding transcription factors such as the p53 tumor suppressor, whose

...
More >>

Cell Biology

Collection of in vivo Capacitated Sperm from Different Locations Along the Reproductive Tract of Time-Mated Female Mice by Microdissection Authors:  Lukas Ded and Jean-Ju Chung, date: 10/20/2021, view: 809, Q&A: 0

Mammalian sperm cells are not capable of fertilizing an egg immediately after ejaculation; instead, they must gradually acquire the capacity to fertilize while they travel inside the female reproductive tract. Sperm cells are transported by the muscular activity of the myometrium to the utero-tubal junction (UTJ) before entering the oviduct where

...
More >>

Developmental Biology

Whole-mount Immunohistochemistry to Visualize Mouse Embryonic Dermal Lymphatic Vasculature Authors:  Xin Geng and R. Sathish Srinivasan, date: 10/20/2021, view: 404, Q&A: 0

Lymphatic vessels are abundant in the skin where they regulate interstitial fluid uptake and immune surveillance. Defects in dermal lymphatic vessels, such as fewer vessels and abnormal lymphatic vessel coverage with mural cells, are frequently associated with lymphedema and other lymphatic disorders. Whole-mount immunohistochemistry allows the

...
More >>
Isolation and Culturing Primary Cholangiocytes from Mouse Liver Authors:  Ramesh Kudira, Bal Krishan Sharma, Mary Mullen, Sujit K. Mohanty, Bryan Donnelly, Gregory M. Tiao and Alexander Miethke, date: 10/20/2021, view: 448, Q&A: 0

Cholangiocytes are epithelial cells lining the intrahepatic and extrahepatic bile ducts. Cholangiocytes perform key physiological functions in the liver. Bile synthesized by hepatocytes is secreted into bile canaliculi, further stored in the gallbladder, and finally discharged into the duodenum. Due to liver injury, biliary epithelial proliferate

...
More >>

Microbiology

A Rapid Induction Overexpression System for the Fission Yeast Schizosaccharomyces pombe Author:  Angad Garg, date: 10/20/2021, view: 306, Q&A: 0

The fission yeast Schizosaccharomyces pombe is an excellent genetically tractable model organism used in the study of conserved eukaryotic cellular biology. One genetic tool in the assessment of gene function is the in vivo overexpression of proteins. Existing overexpression tools have limitations of induction kinetics, dynamic range, and/or

...
More >>
Quantification of RuBisCO Expression and Photosynthetic Oxygen Evolution in Cyanobacteria Authors:  Mateusz Kędzior and Betul Kacar, date: 10/20/2021, view: 422, Q&A: 0

Phototrophic microorganisms are frequently engineered to regulate the expression and the activity of targeted enzymes of interest for specific biotechnological and agricultural applications. This protocol describes a method to evaluate the expression of RuBisCO (ribulose 1,5-bisphosphate carboxylase/oxygenase) in the model cyanobacterium

...
More >>

Neuroscience

Simultaneous Monitoring Cytoplasmic Calcium Ion and Cell Surface Phosphatidylserine in the Necrotic Touch Neurons of Caenorhabditis elegans Authors:  Yoshitaka Furuta and Zheng Zhou, date: 10/20/2021, view: 360, Q&A: 0

Calcium ions trigger many cellular events, including the release of neurotransmitters at the synaptic terminal and excitotoxic cell death. Recently, we have discovered that a transient increase in the level of cytoplasmic Ca2+ triggers the exposure of phosphatidylserine (PS) on the surfaces of necrotic cells in the nematode Caenorhabditis elegans.

...
More >>
Isolation and Electrophysiology of Murine Sympathetic Postganglionic Neurons in the Thoracic Paravertebral Ganglia Authors:  Mallika Halder, Michael Lee McKinnon, Yaqing Li, Peter Wenner and Shawn Hochman, date: 10/20/2021, view: 377, Q&A: 0

The thoracic paravertebral sympathetic chain postganglionic neurons (tSPNs) represent the predominant sympathetic control of vascular function in the trunk and upper extremities. tSPNs cluster to form ganglia linked by an interganglionic nerve and receive multisegmental convergent and divergent synaptic input from cholinergic sympathetic

...
More >>
Isolation and Phospholipid Enrichment of Muscle Mitochondria and Mitoplasts Authors:  Alexandre Prola, Aymeline Vandestienne, Nabil Baroudi, Frederic Joubert, Laurent Tiret and Fanny Pilot-Storck, date: 10/20/2021, view: 333, Q&A: 0

The efficient ATP production in mitochondria relies on the highly specific organization of its double membrane. Notably, the inner mitochondrial membrane (IMM) displays a massive surface extension through its folding into cristae, along which concentrate respiratory complexes and oligomers of the ATP synthase. Evidence has accumulated to highlight

...
More >>

Plant Science

High Throughput Analyses of Ascorbate-turnover Enzyme Activities in Rice (Oryza sativa L.) Seedlings Authors:  Lin-Bo Wu, Yanru Feng, Frederike Zeibig, Muhammad Shahedul Alam and Michael Frei, date: 10/20/2021, view: 509, Q&A: 0

Ascorbate (Vitamin C) fulfills various functions in plant photosynthesis and abiotic stress tolerance. The four key enzymes involved in the ascorbate-turnover pathway are ascorbate peroxidase, ascorbate oxidase, monodehydroascorbate reductase, and dehydroascorbate reductase. Several reports have shown the pivotal roles of these enzymes in plant

...
More >>
Proteoliposomes for Studying Lipid-protein Interactions in Membranes in vitro Author:  Helmut Kirchhoff, date: 10/20/2021, view: 392, Q&A: 0

Lipids in biomembranes can control the structure and, therefore, the functionality of membrane-embedded protein complexes. Unraveling how the lipid composition determines the mode of operation of membrane proteins provides mechanistic insights into their functionality. We applied a proteoliposome technique for studying how proteins function in

...
More >>
High-throughput Screening for Defense Priming-inducing Compounds in Parsley Cell Cultures Authors:  Kathrin Schmitz, Linda Werner and Uwe Conrath, date: 10/20/2021, view: 380, Q&A: 0

Defense priming describes the enhanced potency of cells to activate defense responses. Priming accompanies local and systemic immune responses and can be triggered by microbial infection or upon treatment with certain chemicals. Thus, chemically activating defense priming is promising for biomedicine and agriculture. However, test systems for

...
More >>
We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.