165 Result(s)
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Protocol
Chromatofocusing
Chromatofocusing is a protein-separation technique that was introduced by Sluyterman and his colleagues between 1977 and 1981 (1–5). Chromatofocusing combines the advantage of high-capacity ion-exchange procedure...
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Protocol
I1 PP2A and I2 PP2A
Protein phosphatase 2A (PP2A) is a protein serine/threonine phosphatase that regulates the activities of key rate-limiting enzymes and proteins involved in diverse cellular processes (1–4). However, although much...
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Protocol
Lectin—Gold Histochemistry on Paraffin and Lowicryl K4M Sections Using Biotin and Digoxigenin-Conjugated Lectins
A variety of staining reactions for the visualization of cellular and extracellular glycoconjugates at the light microscopic level are available that are based on the detection of carboxyl and sulfate groups o...
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Protocol
Blot Analysis with Lectins for the Evaluation of Glycoproteins in Cultured Cells and Tissues
The carbohydrate-binding properties of plant and animal lectins have found many diverse applications in biomedical research that are comprehensively treated in the various chapters of this book. In our studies...
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Protocol
Detection of DNA Strand Breakage in the Analysis of Apoptosis and Cell Proliferation by Flow and Laser Scanning Cytometry
The presence of DNA strand breaks resulting from the cleavage of nuclear DNA by the apoptosis-associated endonuclease(s) is one of the most characteristic features of apoptotic cells (1,2). A widely used methodol...
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Protocol
Purification of Proteins Using UltraMacro Spin Columns or ProSorb Sample Preparation Cartridges for Amino Acid Analysis
Amino acid analysis of proteins is one of the best and most accurate methodologies to quantify proteins. Ideally, samples should be pure prior to hydrolysis, however, they are often not only at low concentrati...
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Protocol
Secondary Structure of Peptide Ions in the Gas Phase Evaluated by MIKE Spectrometry
The recent explosive growth of studies of noncovalent interactions of biomolecules using mass spectrometry (1) has greatly stimulated interest in biomolecular conformations in the absence of solvent (2). The two ...
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Protocol
Large Scale Production of Nuclear Receptor Ligand-Binding Domains
The nuclear receptor superfamily is composed of over 150 inducible transcription factors, most of which (1-2) do not have well characterized ligands to date. Nuclear receptors regulate promoters through specific ...
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Protocol
Zymography and Reverse Zymography for Detecting MMPs, andTIMPs
Zymography and reverse zymography are techniques used to analyze the activities of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) in complex biological samples. The two me...
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Protocol
Applications of Constant Denaturant Capillary Electrophoresis and Complementary Procedures
Constant denaturant capillary electrophoresis (CDCE) separates macromolecules based on differences in their melting temperatures. The specific apparatus and operating conditions have been described previously ...
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Protocol
Selective Primer Sequencing from a DNA Mixture by Capillary Electrophoresis
DNA analysis methods have been greatly developed in the past 10 yr with the progress of the Human Genome Project (1). In addition to rapid and high-throughput DNA sequencers (2), various reagents (3), and sample ...
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Protocol
High Resolution Analysis of Point Mutations by Constant Denaturant Capillary Electrophoresis (CDCE)
In recent years, the need for techniques capable of detecting and identifying point mutants has increased dramatically in the fields of genomics, cancer research, and molecular diagnostics. The large arsenal o...
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Protocol
Resolving Racemic Mixtures Using Parallel Combinatorial Libraries
As human enzymes and cell surface receptors possess handedness, the enantiomers of a racemic pair of compounds may be absorbed, activated, and degraded in different manners. In some instances, two enantiomers ...
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Protocol
Structure Determination by NMR
Solution NMR spectroscopy is used widely to determine the structure of proteins. The size of the proteins that can be studied has increased dramatically in the past decade as advances in pulse sequences, probe...
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Protocol
SH3 Domain Protein-Binding Arrays
First identified as part of the Rous sarcoma oncogene product Src, SH3 (Src Homology 3) domains play an important role in intercellular communication and intracellular signal transduction. A high-throughput as...
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Protocol
Liquid Chromatography-Mass Spectrometry and Tandem Mass Spectrometry of Peptides and Proteins
Mass spectrometry has become one of the preferred methods of detection for high-performance liquid chromatography (HPLC) analyses of biopolymers for a broad range of applications. The primary reason for its wi...
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Protocol
Integrated System for Rapid Proteomics Analyses Using Microfluidic Devices Coupled to Nanoelectrospray Mass Spectrometry
This chapter presents an integrated and modular microsystem providing rapid analyses of low femtomole of in-gel digests for proteomics applications. Enhancement of sample throughput is facilitated using an aut...
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Protocol
Intrinsic Fluorescent Detection of Tau Conformation and Aggregation
The polymerization of the microtubule-associated protein tau into paired helical filaments (PHFs) is one of the hallmarks of Alzheimer's disease. Insights into the prerequisites and kinetics of the polymerizat...
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Protocol
Synthesis of Complex Carbohydrates and Glyconjugates
The lipopolysaccharide of capsule-deficient Haemophilus infuenzae strain Rd contains an N-acetylgalactosamine residue attached to the terminal globotriose moiety in the Hex5 glycoform. Genome analysis identified ...
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Protocol
TaqMan® Fluorogenic Detection System to Analyze Gene Transcription in Autopsy Material
Real-time RT-PCR using a TaqMan® fluorogenic detection system is a simple and sensitive assay for quantitative analysis of gene transcription. This method is of potential usefulness in quantifying mRNA of a ta...
