Western blotting

Kurien, Biji T.; Scofield, R. Hal

doi:10.1016/j.ymeth.2005.11.007

Cited by 388 publications

(243 citation statements)

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“…Western blot analysis was performed as previously described (13). In brief, the cells were treated with 0.5 ml radioimmunoprecipitation assay (RIPA) lysis buffer (Beijing ComWin Biotech Co., Ltd., Beijing, China), followed by centrifugation at 16,363 x g at 4˚C for 15 min.…”

Section: Cell Cycle Analysis Cell Cycle Analysis Was Performed Usingmentioning

confidence: 99%

Protective effects of acteoside against X-ray-induced damage in human skin fibroblasts

Yang

Yao

Liu

et al. 2015

Molecular Medicine Reports

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Abstract. To investigate the protective effects of acteoside against apoptosis induced by X-ray radiation in human skin fibroblasts (HSFs), the cells were divided into the following groups: Control group; X-ray radiation group; acteoside group, in which the confluent cells were preincubated with 50 µg/ml acteoside for 2 h followed by radiation; and positive control group, in which the cells were preincubated with 50 µg/ml paeoniflorin followed by radiation. For the radiation, HSF cells preincubated with acteoside or paeoniflorin were exposed to X-ray beams at a dose-rate of 3 Gy/min (16 Gy in total). Cell viability, apoptosis and intracellular alteration of redox were monitored by MTT and flow cytometry. Compared with the radiation group, the number of cells arrested at the G0/G1 phase was significantly reduced in the acteoside and paeoniflorin groups, respectively (P<0.05). X-ray radiation induced marked apoptosis in HSF cells and acteoside reversed this effect. Compared with the radiation group, the generation of intracellular reactive oxygen species (ROS) was abrogated by pre-incubation with acteoside or paeoniflorin (P<0.05). In addition, the upregulation of pro-caspase-3 induced by radiation was reversed by acteoside or paeoniflorin. Radiation could induce upregulation of Bax and downregulation of Bcl-2; however, it was reversed completely after administration of acteoside or paeoniflorin. Furthermore, the enhanced expression of ERK and JNK induced by radiation was reversed by acteoside or paeoniflorin. Acteoside could protect the cells from X-ray induced damage through enhancing the scavenging activity of ROS, decreasing the Bax/Bcl-2 ratio and downregulating the activity of procaspase-3, as well as modulating the mitogen-activated protein kinase signaling pathways.

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Section: Cell Cycle Analysis Cell Cycle Analysis Was Performed Usingmentioning

confidence: 99%

Protective effects of acteoside against X-ray-induced damage in human skin fibroblasts

Yang

Yao

Liu

et al. 2015

Molecular Medicine Reports

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“…The importance of these techniques is reflected in the vast array of literature reporting method development over the last 30 years [5][6][7]. Inquiry-based learning is when students are asked to design and complete experiments where they develop their learning in scientific thinking [8].…”

Section: Discussionmentioning

confidence: 99%

The use of biotin to demonstrate immunohistochemistry, western blotting, and dot blots in university practical classes

Millar

Knighton

Chuck

2012

Biochem Molecular Bio Educ

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Immunological detection of proteins is an essential method to demonstrate to undergraduate biology students, however, is often difficult in resource and time poor student laboratory sessions. This method describes a failsafe method to rapidly and economically demonstrate this technique using biotinylated proteins or biotin itself as targets for detection. This negates the need for a specific primary antibody, saving cost and time. In addition, the easily available and safe reagents allow the methods to be readily adopted without specialist technical expertise. As a result, staff can confidently transfer ownership of the task to the student so as to also develop scientific inquiry skills which promotes student motivation and engagement.

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“…By exposing the blot to an X-ray film, the bands are visualized and further quantitated by densitometry, the results being usually expressed as arbitrary densitometric units. As the method is semi quantitative, it is important that each gel includes samples from all the groups being compared in an experimental design 28,29 .…”

Section: Detecting Brain Proteins: Immunoblot and Proteomicsmentioning

confidence: 99%

Studying the central control of food intake and obesity in rats

Ribeiro

2009

Rev. Nutr.

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A B S T R A C TThe central nervous system regulates energy intake and expenditure through a complex network of neurotransmitters and neuromodulators. It is of great interest to understand the relevance of these systems to the physiological control of energy balance and to the disturbances of obesity. The present paper discusses some of the methods to address this field used at the laboratory of Endocrine Physiology of Universidade Federal de São Paulo. Initially, different experimental models of rat obesity are presented, namely the hypothalamic induced monosodium glutamate model, the Zucker genetic model, and the dietary model. The principles of brain microdialysis are also presented, the technique applied to obtain representative samples of the extracellular fluid of brain sites involved in feeding control. The microdialysate levels of serotonin, an important anorexigenic neurotransmitter, are determined by HPLC with electrochemical detection. The immunoblot technique (Western blot) is used to determine hypothalamic levels of proteins relevant to the anorexigenic effect of serotonin and to analyze the acute activation of the insulin signaling cascade in the hypothalamus. The final section addresses the potential applications of proteomics in the study of the central control of feeding.Indexing terms: Brain microdialysis. Experimental Studies. Food intake. Hypothalamus. Obesity. Rats. R E S U M O

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Western blotting

Cited by 388 publications

References 0 publications

Protective effects of acteoside against X-ray-induced damage in human skin fibroblasts

Protective effects of acteoside against X-ray-induced damage in human skin fibroblasts

The use of biotin to demonstrate immunohistochemistry, western blotting, and dot blots in university practical classes

Studying the central control of food intake and obesity in rats

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