Protein Staining with Ponceau S During Acid-Urea-Triton-Polyacrylamide Gel Electrophoresis

Correia, Heriberto; Rivero, José; Herrera, Flor

doi:10.2144/97225bm12

Cited by 2 publications

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“…Protein size, in kDa, was determined by comparing the relative migration of each band with the banding pattern of standard proteins (Rainbow Markers, Amersham Co., Arlington Heights, IL) of known molecular size: 220, 97, 66, 30, 21 and 14.3 kDa. The separated proteins were then transferred from the PAGE gel to a nitrocellulose (NC) membrane by electroblotting (38), and the protein patterns were visualized by staining with Ponceau S dye (0.3% w/v in 10% acetic acid) (36,39). The protein profiles were developed by washing away excess dye with distilled water, then scanned and saved as digital image files.…”

Section: Methodsmentioning

confidence: 99%

Secretory proteins in the saliva of children

2009

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The protein composition of oral fluid is modulated by environmental factors and physiological states, i.e. chemical, mechanical and pharmacologic stimuli, pathologic conditions, and psychological stress. Secretory protein concentrations in samples of whole saliva (WS) from children were measured and the results were subjected to statistical analysis. Protein expression was determined using electrophoresis and Western blotting. Protein profiles of children were significantly different from those of adults (n = 50, P < 0.05). All samples of saliva from children contained a group of high-molecular-weight (>90 kDa) proteins, whereas fewer than 5% of samples from adults had comparable bands. The ratio of the regulatory subunits (RII) of type II protein kinase A (an enzyme that regulates secretion) to total protein was stable in children's saliva, but variable in saliva from adults. Alpha amylase (alpha-amylase), an enzyme that digests carbohydrates, was less degraded in WS of children than in that of adults. Gingival crevicular fluid of both children and adults did not contain alpha-amylase or RII. No significant gender-based differences were found, but Caucasian children had higher salivary protein levels than children with an African background. Saliva collection is rapid, painless, non-invasive, economical, and yields findings that are reproducible. Objective, biochemical monitoring of secretory proteins in oral fluid of children may reveal responses to stressful stimuli.

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Section: Methodsmentioning

confidence: 99%

Secretory proteins in the saliva of children

2009

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Thyroid hormone receptors α1 and β1 are downregulated in the post-infarcted rat heart: consequences on the response to ischaemia-reperfusion

et al. 2005

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There is accumulating evidence that thyroid hormone metabolism is altered after myocardial infarction (AMI) but its physiological relevance remains largely unknown. The present study investigated the possible role of thyroid hormone signaling in the response of the post-infarcted heart to ischaemia-reperfusion. Wistar rats were subjected to left coronary artery ligation (AMI), or sham operation (SHAM). After 8 weeks, hearts from AMI and SHAM rats were perfused in Langendorff mode and subjected to 20 min of zero-flow global ischaemia (I) and 45 min of reperfusion (R); AMI(I/R), n = 7 and SHAM(I/R), n = 7. Basal left ventricular pressure (LVDP), +dp/dt, and -dp/dt were significantly reduced. Left ventricular weight of the viable myocardium was increased by 14% in the AMI as compared to SHAM hearts, P < 0.05. T(3) and T(4) plasma levels in nM were 1.83 (0.08) and 53.3 (2.9) for SHAM and 1.76 (0.06) and 59.4 (5.2) for AMI rats, respectively, P > 0.05. TRalpha1 and TRbeta1 expression levels were 1.3- and 1.8-fold less in AMI than in SHAM hearts, P < 0.05. Furthermore, SERCA and NHE1 expression levels were 2.1- and 1.8-fold less in AMI than in SHAM, P < 0.05. PKCepsilon was 1.35-fold more in AMI compared to SHAM, P < 0.05. Myocardial glycogen content (in micromol/g) was 7.8 (1.2) in AMI as compared to 4.4 (0.5) for SHAM hearts, P < 0.05. After I/R, left ventricular end-diastolic pressure at 45 min of R (LVEDP45 in mmHg) was 20.3 (3.2) for AMI(I/R) vs 50.6 (4.8) mmHg for SHAM(I/R), P < 0.05. LDH release per gram of tissue was 251 (103) for AMI(I/R) and 762 (74) for SHAM(I/R), P < 0.05. In conclusion, TRalpha1 and TRbeta1 are downregulated after myocardial infarction and this was associated with altered expression of thyroid hormone responsive genes and increased tolerance of the post-infarcted heart to ischaemia-reperfusion injury.

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Protein Staining with Ponceau S During Acid-Urea-Triton-Polyacrylamide Gel Electrophoresis

Cited by 2 publications

References 0 publications

Secretory proteins in the saliva of children

Secretory proteins in the saliva of children

Thyroid hormone receptors α1 and β1 are downregulated in the post-infarcted rat heart: consequences on the response to ischaemia-reperfusion

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