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Article
Interactions between autophagic and endo-lysosomal markers in endothelial cells
Autophagic and endo-lysosomal degradative pathways are essential for cell homeostasis. Availability of reliable tools to interrogate these pathways is critical to unveil their involvement in physiology and pat...
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Article
Open AccessNovel Cell- and Tissue-Based Assays for Detecting Misfolded and Aggregated Protein Accumulation Within Aggresomes and Inclusion Bodies
Aggresomes and related inclusion bodies appear to serve as storage depots for misfolded and aggregated proteins within cells, which can potentially be degraded by the autophagy pathway. A homogenous fluorescen...
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Article
A cell-permeant dye for cell cycle analysis by flow and laser-scanning microplate cytometry
Enzo Life Sciences Nuclear-ID™ Red DNA stain is useful for staging cells on the basis of their DNA content. The stain can be applied to live, detergent-permeabilized or fixed cells for quantification of their dis...
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Protocol
Detection of Glycoproteins in Gels and Blots
As we become more aware of the significance of posttranslational modifications, such as glycosylation, in the production of recombinant proteins and in the proteomic studies of development and disease, techniq...
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Protocol
Background-Free Protein Detection in Polyacrylamide Gels and on Electroblots Using Transition Metal Chelate Stains
Electrophoretically separated proteins may be visualized using organic dyes such as Ponceau Red, Amido Black, Fast Green, or most commonly Coomassie Brilliant Blue (1,2). Alternatively, sensitive detection met...
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Chapter
Deciphering the Hieroglyphics of Functional Proteomics Using Small Molecule Probes
Fluorescence-based total protein detection is generally acknowledged to provide superior capabilities relative to the classical staining methods. Simple measurement of differences in protein expression, howeve...
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Protocol
Quantitative Analysis of Protein Phosphorylation Status and Protein Kinase Activity on Microarrays Using Pro-Q− Diamond Dye Technology
The human genome is estimated to contain 30,000 to 75,000 genes, but as a result of alternative mRNA splicing and protein posttranslational modifications, the human proteome may contain a million or more prote...
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Protocol
Multiplexed Proteomics Technology for the Fluorescence Detection of Phosphorylation and Protein Expression Levels Using Pro-Q® Diamond and SYPRO® Ruby Dyes
The reversible phosphorylation of serine, threonine, and tyrosine residues is arguably one of the most important covalent posttranslational modifications regulating the functional status of proteins in eukaryo...
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Protocol
Sensitive Quantitative Fluorescence Detection of Proteins in Gels Using SYPRO® Ruby Protein Gel Stain
The fluorescent, noncovalent staining method, using SYPRO® Ruby dye resembles colorimetric staining procedures, such as colloidal Coomassie Blue dye staining. The dye provides noncovalent staining and is appli...
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Protocol
Rapid, Sensitive Detection of Proteins in Minigels With Fluorescent Dyes
The advent of polyacrylamide gel electrophoresis (PAGE) in the minigel format (i.e., gel dimensions in the range of 6 × 9 cm or 8 × 8 cm × 0.75 to 1.5 mm thick), the widespread use of precast minigels, and the...
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Protocol
Multiplexed Proteomics Technology for the Fluorescence Detection of Glycoprotein Levels and Protein Expression Levels Using Pro-Q® Emerald and SYPRO® Ruby Dyes
Protein glycosylation is increasingly being recognized as one of the most prominent posttranslational modifications associated with malignant transformation and tumorogenesis as well as cell differentiation. T...
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Chapter
Di- and Tri-Chromatic Fluorescence Detection on Western Blots
The standard protocol for investigators striving to visualize concurrently the total protein profile and a specific antigen by immunodetection is to run replicate gels, using one for general protein staining a...
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Chapter
Multiplexed Proteomics
Two-dimensional gel electrophoresis (2-DGE) has become the central tool for proteome analysis and especially for the comparison of changes in protein expression levels arising from disease progression or thera...
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Protocol
Detection of Glycoproteins in Gels and Blots
As we become more aware of the significance of posttranslational modifications, such as glycosylation, in the production of recombinant proteins and in the proteomic studies of development and disease, techniq...
-
Protocol
Background-Free Protein Detection in Polyacrylamide Gels and on Electroblots Using Transition Metal Chelate Stains
Electrophoretically separated proteins may be visualized using organic dyes such as Ponceau Red, Amido Black, Fast Green, or most commonly Coomassie Brilliant Blue (1,2). Alternatively, sensitive detection method...
-
Protocol
Protein Detection Using Reversible Metal Chelate Stains
Electrophoretically separated proteins may be visualized using organic dyes, such as Ponceau red, Amido black, fast green, or most commonly Coomassie Brilliant Blue (1,2). Alternatively, sensitive detection metho...
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Protocol
Image Acquisition in 2-D Electrophoresis
Prevalent methods for visualizing proteins resolved by two-dimensional (2-D) gel electrophoresis include autoradiography, silver staining, and Coomassie brilliant blue staining (1,2. The organic dye Coomassie bri...
