A simplified method for the quantitative assay of small amounts of protein in biologic material

Schacterle, George R.; Pollack, Robert L.

doi:10.1016/0003-2697(73)90523-x

Cited by 1,752 publications

(482 citation statements)

References 2 publications

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“…Specific activity is expressed as/amoles of p-nitrophenol released min-lmg -1 of protein. Protein concentration was determined according to Schacterle and Pollack [ 12].…”

Section: Methodsmentioning

confidence: 99%

Alkaline phosphatase activity associated to a calcium binding glycoprotein from calf scapula cartilage

Vittur

Bernard

1973

FEBS Letters

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“…Specific activity is expressed as/amoles of p-nitrophenol released min-lmg -1 of protein. Protein concentration was determined according to Schacterle and Pollack [ 12].…”

Section: Methodsmentioning

confidence: 99%

Alkaline phosphatase activity associated to a calcium binding glycoprotein from calf scapula cartilage

Vittur

Bernard

1973

FEBS Letters

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“…Samples were centrifuged at 13,000 g at 4°C for 5 min to remove insoluble material. The protein content of the clarified extracts was determined by a modification of the Lowry method [15]. Samples were diluted to a common concentration of 0.4 mg/ml in Laemmli buffer and 10 μg total protein subjected to SDS-PAGE.…”

Section: Muscle Biopsymentioning

confidence: 99%

Low birthweight is associated with specific changes in muscle insulin-signalling protein expression

et al. 2005

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Aims/hypothesis: People with low birthweight have an increased risk of developing type 2 diabetes mellitus in adulthood. The mechanistic basis of this phenomenon is not known. Here we investigate the effect of early growth restriction on the expression of insulin-signalling proteins in skeletal muscle in a human cohort and a rat model. Methods: We recruited 20 young men with low birthweight (mean birthweight 2702±202 g) and 20 age-matched control subjects (mean birthweight 3801± 99 g). Biopsies were obtained from the vastus lateralis muscle and protein expression of selected insulin-signalling proteins was determined. Rats used for this study were male offspring born to dams fed a standard (20%) protein diet or a low (8%) protein diet during pregnancy and lactation. Protein expression was determined in soleus muscle from adult offspring. Results: Low-birthweight subjects showed reduced muscle expression of protein kinase C (PKC)ζ, p85α, p110β and GLUT4. PKCζ, GLUT4 and p85 were also reduced in the muscle of rats fed a low-protein diet. Other proteins studied were unchanged in low-birthweight humans and in rats fed a low-protein diet when compared with control groups. Conclusions/interpretation: We found decreased expression of specific insulin-signalling proteins in low-birthweight subjects compared to controls. These changes precede the onset of impaired glucose tolerance. The similarity of protein expression profile in the men with low birthweight compared to that of the offspring of rats fed a low-protein diet suggests that the rodent model is an accurate representation of the human situation. It also provides a potential mechanistic explanation as to why the fetal environment plays an important role in determining risk of developing type 2 diabetes.

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“…The LDL was dialysed against 3 × 4 litres 140 mM NaCl, 8.1 mM Na 2 HPO 4 , 1.9 mM NaH 2 PO 4 , pH 7.4, containing 100 µM EDTA, sterilised by membrane filtration and assayed for protein by a modified Lowry assay [57]. Separate LDL preparations were used to repeat the experiments.…”

Section: Isolation Of Ldlmentioning

confidence: 99%

Mechanism of the antioxidant to pro-oxidant switch in the behavior of dehydroascorbate during LDL oxidation by copper(II) ions

Horsley

Burkitt

Jones

et al. 2007

Archives of Biochemistry and Biophysics

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Subject area: Systems biochemistryAbbreviations: BCDS, bathocuproine disulphonic acid; DMPO,5, EPR, electron paramagnetic resonance; 13-HPODE, 13(S)-hydroperoxyoctadeca-9Z,11E-dienoic acid 2 ABSTRACTOxidised low density lipoprotein (LDL) may be involved in the pathogenesis of atherosclerosis.We have therefore investigated the mechanisms underlying the antioxidant/pro-oxidant behavior of dehydroascorbate, the oxidation product of ascorbic acid, toward LDL incubated with Cu 2+ ions. By monitoring lipid peroxidation through the formation of conjugated dienes and lipid hydroperoxides, we show that the pro-oxidant activity of dehydroascorbate is critically dependent on the presence of lipid hydroperoxides, which accumulate during the early stages of oxidation.Using electron paramagnetic resonance spectroscopy, we show that dehydroascorbate amplifies the generation of alkoxyl radicals during the interaction of copper ions with the model alkyl hydroperoxide, tert-butylhydroperoxide. Under continuous-flow conditions, a prominent doublet signal was detected, which we attribute to both the erythroascorbate and ascorbate free radicals.On this basis, we propose that the pro-oxidant activity of dehydroascorbate toward LDL is due to its known spontaneous interconversion to erythroascorbate and ascorbate, which reduce Cu 2+ to Cu + and thereby promote the decomposition of lipid hydroperoxides. Various mechanisms, including copper chelation and Cu + oxidation, are suggested to underlie the antioxidant behavior of dehydroascorbate in LDL that is essentially free of lipid hydroperoxides.

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A simplified method for the quantitative assay of small amounts of protein in biologic material

Cited by 1,752 publications

References 2 publications

Alkaline phosphatase activity associated to a calcium binding glycoprotein from calf scapula cartilage

Alkaline phosphatase activity associated to a calcium binding glycoprotein from calf scapula cartilage

Low birthweight is associated with specific changes in muscle insulin-signalling protein expression

Mechanism of the antioxidant to pro-oxidant switch in the behavior of dehydroascorbate during LDL oxidation by copper(II) ions

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