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Developmental Biology

Phalloidin Staining and Immunohistochemistry of Zebrafish Embryos Authors:  Michelle F. Goody and Clarissa A. Henry, date: 06/05/2013, view: 13498, Q&A: 1
Fluorescent conjugated Phalloidin is a stain that allows for visualization of F-actin. In immunohistochemistry, primary antibodies and fluorescent conjugated secondary antibodies can be used to visualize subcellular localization and relative amounts of proteins of interest. Here is a protocol for Phalloidin and antibody staining of zebrafish ...
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Motility Assay for Zebrafish Embryos Authors:  Michelle F. Goody and Clarissa A. Henry, date: 06/05/2013, view: 5802, Q&A: 0
Analyzing the swimming ability of 2 days post fertilization zebrafish embryos can be a useful technique to study neuromuscular function. Here is a protocol for determining the time it takes for zebrafish embryos to swim a predetermined distance.
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Immunology

Generation of Polyclonal Specific Antibodies Authors:  Maureen Wirschell and Mary E. Porter, date: 06/05/2013, view: 4870, Q&A: 0
Generation of antibodies specific for a protein of interest is a common method in many disciplines. This protocol details the steps in production of a polyclonal antibody in rabbits using a bacterially expressed fusion protein as an antigen. The protocol is generated based on data presented in Wirschell et al.(2013).
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Microbiology

Quantification of Retro- and Lentiviral Reverse Transcriptase Activity by Real-time PCR Authors:  Jolien Vermeire and Bruno Verhasselt, date: 06/05/2013, view: 5445, Q&A: 0
Quantification of retroviral reverse transcriptase activity in retrovirus containing supernatant by quantitative reverse transcription PCR as a method for titration of HIV, lenti- and retroviral vectors is described here.. The procedure was optimized for use with LightCycler 480 (Roche, Vilvoorde, Belgium) and ABI 7300 real-time PCR system ...
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Detection of Membrane Potential in Mycobacterium tuberculosis Authors:  Manbeena Chawla and Amit Singh, date: 06/05/2013, view: 6310, Q&A: 2
DiOC2 (Novo et al., 2000) exhibits green fluorescence in all bacterial cells, but the fluorescence shifts towards red emission as the dye molecules self associate at the higher cytosolic concentrations caused by larger membrane potentials. Proton ionophores such as CCCP destroy membrane potential by eliminating the proton ...
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Neuroscience

Electroolfactogram (EOG) Recording in the Mouse Main Olfactory Epithelium Authors:  Xuanmao Chen, Zhengui Xia and Daniel R. Storm, date: 06/05/2013, view: 4183, Q&A: 0
Olfactory sensory neurons in the main olfactory epithelium (MOE) are responsible for detecting odorants and EOG recording is a reliable approach to analyze the peripheral olfactory function. However, recently we revealed that rodent MOE can also detect the air pressure caused by airflow. The sensation of airflow pressure and odorants may function ...
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Plant Science

2D Diagonal Redox SDS-PAGE of Proteins Authors:  Christian Schwarz and Joerg Nickelsen, date: 06/05/2013, view: 7023, Q&A: 0
2D diagonal redox SDS-PAGE of proteins is used to detect intramolecular or intermolecular disulfide bridges using Chlamydomonas in this example (Stroeher and Dietz, 2008; Schwarz et al., 2012). Both dimensions consist of a conventional SDS-PAGE, except that the sample buffer for the first dimension lacks a reducing agent. ...
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Wax Analysis of Stem and Rosette Leaves in Arabidopsis thaliana Authors:  Tegan M Haslam and Ljerka Kunst, date: 06/05/2013, view: 6111, Q&A: 1
The primary aerial surfaces of all land plants are coated by a lipidic cuticle, which restricts non-stomatal water loss and protects the plant from pathogens, herbivores, and ultraviolet radiation. The cuticle is made up of two components: cutin, a polymer of hydroxy- and epoxy- long-chain fatty acid derivatives and glycerol, and cuticular waxes, ...
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35S pulse Labelling of Chlamydomonas Chloroplast Proteins Authors:  Alexandra-Viola Bohne, Christian Schwarz and Joerg Nickelsen, date: 06/05/2013, view: 4993, Q&A: 0
35S pulse labelling of proteins is used to attach a radioactive label to newly synthesized proteins, as sulfur is an element that is mainly present in proteins (Fleischmann and Rochaix 1999). Depending on your organism’s uptake mechanisms you need cysteine, methionine or sulfuric acid as a source of radioactive sulfur. This example uses ...
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Systems Biology

Detection of DNA Methylation Changes Surrounding Transposable Elements Authors:  Beery Yaakov and Khalil Kashkush, date: 06/05/2013, view: 5249, Q&A: 0
Transposable elements (TEs) are a major component of all genomes, thus the epigenetic mechanisms controlling their activity is an important field of study. Cytosine methylation is one of the factors regulating the transcription and transposition of TEs, alongside Histone modifications and small RNAs. Adapter PCR-based methods [such as Amplified ...
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High Resolution Detection of Genetic Changes Associated with Transposons Authors:  Beery Yaakov and Khalil Kashkush, date: 06/05/2013, view: 4426, Q&A: 0
Transposable elements (TEs) are repetitive sequences, capable of inducing genetic mutations through their transpositional activity, or by non-homologous or illegitimate recombination. Because of their similarity and often high copy numbers, examining the effects of mutations caused by TEs in different samples (tissues, individuals, species, etc.) ...
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