Selective silver-staining methods for RNA and proteins in the same polyacrylamide gels

Paleologue, Anne; Reboud, Jean‐Paul; Reboud, Anne-Marie

doi:10.1016/0003-2697(88)90279-5

Cited by 12 publications

(2 citation statements)

References 13 publications

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“…The LDL was aspirated from the gradient between densities of 1.030 and 1.050 g/ml, adjusted with KBr to a density of 1.21 g/ml, layered under a KBr solution in water, of the same density, and ultracentrifuged to purify further and to concentrate LDL. SDS/PAGE of the LDL showed that apo B1OO was the only protein present in the isolated LDL, as determined by silver staining as described by Paleologue et al [30]. SDS/PAGE was performed under reducing conditions by the method of Laemmli [31], with resolving gels containing a gradient of 4-20 % (w/v) acrylamide and stacking gels of 3.5 % (w/v) acrylamide.…”

Section: Cell Culturementioning

confidence: 99%

Butyrate stimulates the secretion of apolipoprotein (apo) A-I and apo B100 by the human hepatoma cell line Hep G2. Induction of apo A-I mRNA with no change of apo B100 mRNA

Kaptein

Roodenburg

Princen

1991

Biochemical Journal

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Addition of sodium butyrate to the culture medium of the human hepatoma cell line Hep G2 resulted in a time- and dose-dependent increase in the secretion of apolipoprotein A-I (apo A-I) and apolipoprotein B100 (apo B100). After a 24 h preincubation period, a 2.4- and 2.2-fold increase in the secretion of apo A-I and apo B100 respectively was obtained during the next 24 h in the presence of 2 mM-sodium butyrate. Secretion of albumin, fibrinogen or [35S]methionine-labelled newly synthesized proteins was unaffected or only marginally affected, indicating that the effect of butyrate on apo A-I and apo B100 is not part of a general effect on protein synthesis and secretion. In structure-function studies, butyrate was found to be the most potent inducer among various straight-chain carboxylic acids. Hydroxylated, aminated and otherwise modified butyrate derivatives were inactive. The enhanced accumulation of apo A-I and apo B100 in the culture medium could not be explained by changes in the uptake and degradation of the synthesized apolipoproteins or by alterations in the secretion of possible intracellular pools. In addition, [35S]methionine incorporation studies indicated that synthesis and/or secretion of newly synthesized apo A-I and apo B100 is enhanced in the presence of butyrate. The apo A-I mRNA level was increased 2.3-fold upon treatment with 2 mM-butyrate for 48 h, suggesting regulation at (post-)transcriptional level. In contrast, no change in the level of apo B100 mRNA in butyrate-treated cells was observed, indicating regulation at translational or co- or post-translational level. We propose that the effect of butyrate on the secretion of apo A-I and apo B100 by Hep G2 results from two different regulatory mechanisms.

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Section: Cell Culturementioning

confidence: 99%

Butyrate stimulates the secretion of apolipoprotein (apo) A-I and apo B100 by the human hepatoma cell line Hep G2. Induction of apo A-I mRNA with no change of apo B100 mRNA

Kaptein

Roodenburg

Princen

1991

Biochemical Journal

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“…The silver diammine method remains popular and has been used recently for demonstrating neuronal bodies, 69,70 fungi, 71 and glomerular basement membrane. 72 It has also been used in combination with microwaves 72,73 and for visualizing proteins and other substances on gels [74][75][76][77][78] and diaminobenzidine (DAB) in immunocytochemistry. 79 An alternative, the 'protargol' or silver proteinate method, allegedly gives a cleaner background in tissue sections.…”

Section: Introductionmentioning

confidence: 99%

Silver development in microscopy and bioanalysis: past and present

Newman

Jasani

1998

J. Pathol.

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Quantitative proteomics: assessing the spectrum of in-gel protein detection methods

2010

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Proteomics research relies heavily on visualization methods for detection of proteins separated by polyacrylamide gel electrophoresis. Commonly used staining approaches involve colorimetric dyes such as Coomassie Brilliant Blue, fluorescent dyes including Sypro Ruby, newly developed reactive fluorophores, as well as a plethora of others. The most desired characteristic in selecting one stain over another is sensitivity, but this is far from the only important parameter. This review evaluates protein detection methods in terms of their quantitative attributes, including limit of detection (i.e., sensitivity), linear dynamic range, inter-protein variability, capacity for spot detection after 2D gel electrophoresis, and compatibility with subsequent mass spectrometric analyses. Unfortunately, many of these quantitative criteria are not routinely or consistently addressed by most of the studies published to date. We would urge more rigorous routine characterization of stains and detection methodologies as a critical approach to systematically improving these critically important tools for quantitative proteomics. In addition, substantial improvements in detection technology, particularly over the last decade or so, emphasize the need to consider renewed characterization of existing stains; the quantitative stains we need, or at least the chemistries required for their future development, may well already exist.

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Selective silver-staining methods for RNA and proteins in the same polyacrylamide gels

Cited by 12 publications

References 13 publications

Butyrate stimulates the secretion of apolipoprotein (apo) A-I and apo B100 by the human hepatoma cell line Hep G2. Induction of apo A-I mRNA with no change of apo B100 mRNA

Butyrate stimulates the secretion of apolipoprotein (apo) A-I and apo B100 by the human hepatoma cell line Hep G2. Induction of apo A-I mRNA with no change of apo B100 mRNA

Silver development in microscopy and bioanalysis: past and present

Quantitative proteomics: assessing the spectrum of in-gel protein detection methods

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