An evaluation of the Coomassie brilliant blue G-250 dye-binding method for quantitative protein determination

Pierce, James C.; Suelter, Clarence H.

doi:10.1016/0003-2697(77)90723-0

Cited by 254 publications

(70 citation statements)

References 9 publications

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“…After passage through a 0.45-,um filter, the cell-free lavage supernates were concentrated by pressure dialysis to 2-3 ml over a YM5 membrane (Amicon Corp., Lexington, MA) and stored at -80°C. (21,22). These protein-specific sensitivity differences are substantially diminished when sodium dodecyl sulfate (SDS) is added to the reaction medium.…”

Section: Methodsmentioning

confidence: 99%

Protein permeability in the adult respiratory distress syndrome. Loss of size selectivity of the alveolar epithelium.

Holter¹,

Weiland²,

Pacht³

et al. 1986

J. Clin. Invest.

220

108

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Small amounts of plasma protein normally reach the alveolar epithelial surface by a size-selective process that restricts the passage of very large molecules. Size selectivity may be compromised in the lungs of patients with the adult respiratory distress syndrome (ARDS). To assess this question, bronchoalveolar lavage fluid (BALF) from normal volunteers (n 11), cardiac edema patients (n = 3), and ARDS patients (a = 8) was compared. Mean total protein in ARDS BALF was >12 times the levels in normals or cardiac edema patients. BALF/plasma total protein ratios and measurements of epithelial lining fluid protein also separated the patient groups. The large proteins IgM and a2-macroglobulin were found in ARDS BALF at >90 times the concentrations of normal or cardiac edema fluid. The relationship of distribution coefficient vs. log molecular weight for seven proteins (54,000-900,000 mol wt) hyperbolically increased in normals but was flat in ARDS patients. Sodium dodecyl sulfatepolyacrylamide gel electrophoresis revealed a paucity of high molecular weight proteins in normal and cardiac edema BALF, but demonstrated the full spectrum of plasma proteins in ARDS BALF. We conclude that normal size selectivity is preserved in cardiac edema but is destroyed by the alveolar-capillary injury of ARDS.

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Section: Methodsmentioning

confidence: 99%

Protein permeability in the adult respiratory distress syndrome. Loss of size selectivity of the alveolar epithelium.

Holter¹,

Weiland²,

Pacht³

et al. 1986

J. Clin. Invest.

220

108

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“…The activity of aminopeptidase N (APN) was determined according to Maroux et al (1973). The method described by Pierce and Suelter (1977) was used to estimate the protein concentration of mucosa samples.…”

Section: Small Intestinal Enzyme Activities Determinationmentioning

confidence: 99%

Adaptability of the digestive function according to age at weaning in the rabbit: I. Effect on feed intake and digestive functionality

Gallois

Huërou‐Luron

Fortun-Lamothe

et al. 2008

Animal

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The functional adaptability of the digestive system to the level of feed intake was investigated in the young rabbits by comparing two groups of 12 litters each, weaned at 21 (W21) or 35 (W35) days of age. From 14 days onwards, rabbits were fed a pelleted feed (NDF: 332 g/kg, CP: 177 g/kg, starch: 98 g/kg, as-fed basis). Until 49 days of age, the profile of digestive enzymes was weekly determined in the small intestinal content and mucosa, as well as caecal fermentation traits and fibrolytic activities. In the W21 group, the solid feed intake was increased by 57% between 21 and 35 days (P , 0.01), while the daily body growth was lower from 21 till 42 days (217%, P , 0.05) when compared with the W35 group. Activities of enzymes of pancreatic origin were only scarcely influenced by the weaning age. In the W21 group, amylase activity tended to be lower at 28 days of age (236%, P 5 0.064), and trypsin activity was decreased by 31% at 49 days of age (P , 0.01). Lipase activity was similar in both weaning groups. Duodenal and jejunal activities of maltase and aminopeptidase N (APN) were higher on day 28 in the W21 group as compared with the W35 group (31.4 to 32.4, respectively, P , 0.05). On day 35, duodenal APN activity was twice as higher in the W21 group than in the W35 group (P , 0.01). In caecum, major differences between both weaning groups were observed at 28 days of age with a decrease in ammonia concentration (243%, P , 0.01) in W21 compared with W35 rabbits. Conversely, the acetate proportion was 5% higher in the W21 group (P , 0.01) on day 28. In conclusion, the digestive tract of early-weaned rabbits showed some adaptative properties in response to nutritional environment changes, but they were insufficient to maintain their growth rate.

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“…Enzyme and substrate controls were also performed simultaneously by replacing substrate and enzymes by the MES buffer, respectively. The protein content of the enzyme preparations was determined according to Pierce and Suelter [20] using bovine serum albumin as the standard. Enzyme activities were expressed as the quantity of reducing sugar (for polysaccharidases) or p-nitrophenol (for glycosidases) released per mg protein per 1 h for specific activity or g DM solid phase per 1 h for total activity.…”

Section: Enzyme Preparation and Fibrolytic Activity Measurementmentioning

confidence: 99%

Cereal supplementation modified the fibrolytic activity but not the structure of the cellulolytic bacterial community associated with rumen solid digesta

et al. 2001

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-4 ruminally cannulated cows were fed a forage diet (93% hay + 7% straw) and a mixed diet (33 % hay + 7% straw + 40% barley) in a 2 ¥ 2 crossover experimental design. In sacco degradation of forage, fibrolytic activities (polysaccharidases and glycosidases) of the solid-associated bacteria (SAB), and distribution of the 3 main cellulolytic bacterial species (Fibrobacter succinogenes, Ruminococcus albus, Ruminococcus flavefaciens) were determined for both diets. Barley supplementation decreased the hay degradation rate and mainly the polysaccharidase activities of the SAB (30% on average). The sum of rRNA of the 3 cellulolytic bacterial species represented on average 17% of the total bacterial signal and R. albus was the dominant cellulolytic bacterial species of the 3 studied. Barley supplementation did not modify the proportion of the 3 cellulolytic bacteria attached to plant particles. The negative effect of barley on the ruminal hay degradation rate is due to a decrease in fibrolytic activity of the SAB, and not to a modification of the balance of the three cellulolytic bacterial species examined. fibrolytic activity / oligonucleotide probe / cellulolytic bacteria / cereal supplementation / rumen Résumé -La supplémentation en céréale modifie l'activité fibrolytique mais pas la structure de la communauté bactérienne cellulolytique associée à la phase solide du contenu ruminal. 4 vaches, canulées du rumen, ont été alimentées avec un régime fourrage (93 % foin + 7 % paille) et un régime mixte (33 % foin + 7 % paille + 60 % orge) dans un schéma croisé 2 ¥ 2. La dégradation Reprod. Nutr. Dev. 41 (2001) 413-424 413

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An evaluation of the Coomassie brilliant blue G-250 dye-binding method for quantitative protein determination

Cited by 254 publications

References 9 publications

Protein permeability in the adult respiratory distress syndrome. Loss of size selectivity of the alveolar epithelium.

Protein permeability in the adult respiratory distress syndrome. Loss of size selectivity of the alveolar epithelium.

Adaptability of the digestive function according to age at weaning in the rabbit: I. Effect on feed intake and digestive functionality

Cereal supplementation modified the fibrolytic activity but not the structure of the cellulolytic bacterial community associated with rumen solid digesta

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