The Bradford Method For Protein Quantitation

Kruger, Nicholas J.

doi:10.1007/978-1-59745-198-7_4

Cited by 579 publications

(309 citation statements)

References 16 publications

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“…Samples (50 mg DW) for protein analysis were digested in 1 N NaOH for 24 h at room temperature and quantified according to Kruger (Kruger 1994), using bovine serum albumin as the standard.…”

Section: Total Protein Analysismentioning

confidence: 99%

Growth, protein and carbohydrate contents in Ulva rigida and Gracilaria bursa-pastoris integrated with an offshore fish farm

2015

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Low-technology practices are generally the rule when cultivating marine macroalgae, and they do not necessarily comply with sustainability requirements. When integrated with other marine organisms in land-based setups, seaweed culture can be sustainable also providing environmental benefits. Major challenges of such integrated aquaculture systems, however, are in sea-based setups. The current study examined the growth rates of Ulva rigida and Gracilaria bursa-pastoris, as well as their protein and carbohydrate contents, when exposed to different distances from an offshore, fed-fish cage system. Nutrient levels in seawater were consistently high downstream from the fish cages, significantly enhancing the specific growth rates and cellular contents of starch and soluble protein in these two seaweeds. Specifically, daily maximal growth rates were 17 % day −1 for U. rigida and 10 % day −1 for G. bursa-pastoris, maximal starch contents were 22 and 21 %, respectively, and maximal protein contents were on a dry weight basis 7 and 13 %, respectively. When repositioned at low ambient nutrient levels for 48 h, the starch and the carbohydrate levels increased by 129 and 131 % for U. rigida and by 198 and 150 % for G. bursa-pastoris, respectively. Altogether, this study supports implementing future viable mean of sustainable macroalgae cultivation by taking advantage of excessive nutrients released from an offshore fish farm.

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Section: Total Protein Analysismentioning

confidence: 99%

Growth, protein and carbohydrate contents in Ulva rigida and Gracilaria bursa-pastoris integrated with an offshore fish farm

2015

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“…1 9 10 5 cells were treated 4 h with iron in concentration 250 lM or IL1b in concentration 10 ng/ml or both iron and IL1b. The values are means of triplicate cultures ± SE Cutting Edge in Autoimmunity content was determined by the Bradford method[15] using the Bradford Ultra Assay (Expedeon).Briefly, for FPN and TfR, 150 lg samples were electrophoresed on a 12 % SDS-polyacrylamide gel. For L-and Hferritin, samples were loaded on a 15 % SDS-polyacrylamide gel.…”

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confidence: 99%

β-Carotene can reverse dysregulation of iron protein in an in vitro model of inflammation

2014

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Anemia of chronic disease is frequently seen in chronic inflammatory conditions. Its hallmark is disrupted iron homeostasis, with increased uptake and retention of iron in cells of the reticuloendothelial system. Using the Caco-2 cell line as an in vitro model for iron absorption, local intestinal iron-related protein dynamics were evaluated during interleukin (IL)1β/iron-induced inflammation, confirmed by IL8 release, and following β-carotene and vitamin A supplementation. Time- and dose-dependent iron administration to the cells was then studied. The effects on heavy and light ferritin, ferroportin, transferrin receptor and intracellular iron levels were compared in inflamed Caco-2 cells with and without application of the anti-inflammatory agents β-carotene and vitamin A. IL1β treatment led to IL8 release, a surge in both ferritins' expressions and suppression of ferroportin and transferrin receptor expression. β-Carotene significantly reduced IL8 (1,306.2-253.75 pg/ml), decreased light and heavy ferritin by 77.8 and 45.8%, respectively, and increased ferroportin by 59.9% (P < 0.05). Increasing iron concentrations and incubation periods resulted in increased IL8 release. A strong correlation was found between the levels of IL8 and the ferritins. Intracellular iron sequestration was induced by IL1β and iron and alleviated by β-carotene. β-Carotene normalized the main iron-related proteins' levels, reduced IL8 production, and released intracellular trapped iron. These results highlight local mucosal control of iron regulation and suggest that by applying anti-inflammatory compounds, less iron is locked in inflamed intestinal epithelial cells, leading to its increased bioavailability. This suggests a possible approach to combating anemia associated with chronic inflammatory conditions.

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“…Bradford assay was used to measure protein concentration (Kruger 1994). Then protein samples were electrophoresed on 12 % SDS-polyacrylamide gel (SDS-PAGE) and transferred to polyvinylidene fluoride (PVDF) membranes.…”

Section: Western Blot Analysismentioning

confidence: 99%

Gas1 Knockdown Increases the Neuroprotective Effect of Glial Cell-Derived Neurotrophic Factor Against Glutamate-Induced Cell Injury in Human SH-SY5Y Neuroblastoma Cells

et al. 2015

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Growth arrest-specific 1 (Gas1) protein acts as an inhibitor of cell growth and a mediator of cell death in nervous system during development and is also re-expressed in adult neurons during excitotoxic insult. Due to its structural similarity to the glial cell-derived neurotrophic factor family receptors α (GFRα), Gas1 is likely to interfere with the neuroprotective effect of GDNF. In the present study, we investigated the expression profile of Gas1 during glutamate insults in human SH-SY5Y neuroblastoma cells as well as the influence of Gas1 inhibition on the protective effect of GDNF against glutamate-induced cell injury. Our data showed that Gas1 expression was significantly increased with the treatment of glutamate in SH-SY5Y cells. The silencing of Gas1 by small interfering RNA promoted the protective effect of GDNF against glutamate-induced cytotoxicity as well as cell apoptosis, which effect was likely mediated through activating Akt/PI3 K-dependent cell survival signaling pathway and inhibiting mitochondrial-dependent cell apoptosis signaling pathway via Bad dephosphorylation blockade. In summary, this study showed the synergistic effect of Gas1 inhibition and GDNF against glutamate-induced cell injury in human SH-SY5Y neuroblastoma cells, which information might significantly contribute to better understanding the function of Gas1 in neuronal cells and form the basis of the therapeutic development of GDNF in treating human neurodegenerative diseases in the future.

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The Bradford Method For Protein Quantitation

Cited by 579 publications

References 16 publications

Growth, protein and carbohydrate contents in Ulva rigida and Gracilaria bursa-pastoris integrated with an offshore fish farm

Growth, protein and carbohydrate contents in Ulva rigida and Gracilaria bursa-pastoris integrated with an offshore fish farm

β-Carotene can reverse dysregulation of iron protein in an in vitro model of inflammation

Gas1 Knockdown Increases the Neuroprotective Effect of Glial Cell-Derived Neurotrophic Factor Against Glutamate-Induced Cell Injury in Human SH-SY5Y Neuroblastoma Cells

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