Two-dimensional phos-tag zymograms for tracing phosphoproteins by activity in-gel staining
نویسندگان
چکیده
منابع مشابه
Two-dimensional phos-tag zymograms for tracing phosphoproteins by activity in-gel staining
Protein phosphorylation is one of the most common post-translational modifications regulating many cellular processes. The phos-tag technology was combined with two-dimensional zymograms, which consisted of non-reducing IEF PAGE or NEPHGE in the first dimension and high resolution clear native electrophoresis (hrCNE) in the second dimension. The combination of these electrophoresis methods was ...
متن کاملQuantitative detection of phosphoproteins by combination of two-dimensional difference gel electrophoresis and phosphospecific fluorescent staining.
Here we combine a standard two-dimensional difference gel electrophoresis (DIGE) protocol with subsequent post-staining of gels with phosphospecific fluorescent Pro-Q Diamond dye. The combination of these two methods for fluorescence detection of proteins allows quantitative detection of phosphoproteins in 2-DE-gels. We established this protocol within a functional proteomics experiment. Mammar...
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The low specificity of anti-phosphoprotein antibodies is often a problem in immunoblotting analyses. We introduce a simple pretreatment procedure for cell lysates to give more specific detection of phosphoproteins in immunoblotting. Cellular phosphoproteins were preferentially trapped on Phos-tag agarose phosphate-affinity beads in a homemade spin-centrifuge microtube unit, and nonphosphorylate...
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Pro-Q Diamond (Pro-Q DPS) is a commercially available stain that binds the phosphate moiety of phosphoproteins with high sensitivity and linearity. To conserve consumable costs we demonstrate that threefold diluted Pro-Q DPS offers the same sensitivity and linearity of signal to that obtained with undiluted Pro-Q DPS. The optimal conditions for Pro-Q DPS indicate that fixation, staining, and de...
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The principal phosphoglycoproteins of avian and amphibian egg yolk, known as phosvitins, have been examined extensively in developmental studies of estrogenic induction of protein synthesis (1) and, more recently, as substrates for protein kinases which phosphorylate the basic nuclear proteins (2,3). Phosvitin has few aromatic and basic amino acids (4) which can be detected by uv absorbance or ...
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ژورنال
عنوان ژورنال: Frontiers in Plant Science
سال: 2015
ISSN: 1664-462X
DOI: 10.3389/fpls.2015.00230