Removal of lipid contaminants by organic solvents from oilseed protein extract prior to electrophoresis

Wang, Wei; Vignani, Rita; Scali, Monica; Sensi, Elisabetta; Tiberi, P.; Cresti, Mauro

doi:10.1016/j.ab.2004.02.044

Cited by 39 publications

(30 citation statements)

References 10 publications

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“…Afterwards, lipids dissolved in the chloroform phase can be further removed by aqueous TCA washes. The protocol was found to work well in other oil crop seeds such as olive, sunflower, rapeseed, and sesame [67].…”

Section: Castor Seedmentioning

confidence: 94%

“…When protein extracts of castor seeds are resolved by electrophoresis, the resolution of proteins is poor mainly due to the interference of lipids. Wang et al [67] developed a protocol based on the chloroform/TCA/acetone precipitation protocol for the removal of excess lipids from oil seed extracts prior to 2-DE. In the protocol, protein extract of dry castor seeds was mixed well with an equal volume of chloroform/methanol (2:1).…”

Section: Castor Seedmentioning

confidence: 99%

See 1 more Smart Citation

Optimizing protein extraction from plant tissues for enhanced proteomics analysis

Wang¹,

Tai

Chen

2008

J of Separation Science

Self Cite

148

110

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Plant tissues usually contain high levels of proteases and secondary metabolites that severely interfere with protein extraction, separation, and identification. Preparation of high-quality protein samples from plant tissues for proteomic analysis represents a great challenge. This article briefly describes the critical points in protein separation, especially secondary metabolites in plant tissues, and removal strategy. It provides an updated overview of three total protein extraction methods and their applications in proteomic analysis of various recalcitrant tissues.

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Section: Castor Seedmentioning

confidence: 94%

Section: Castor Seedmentioning

confidence: 99%

Optimizing protein extraction from plant tissues for enhanced proteomics analysis

Wang¹,

Tai

Chen

2008

J of Separation Science

Self Cite

148

110

View full text Add to dashboard Cite

show abstract

“…One of the studies in the method development of protein extraction from plants was carried out using TCA/acetone method (Wang et al, 2004). This method was also employed by Saravanan and Rose (2004) which noticed missing proteins, possibly due to protein solubility issues.…”

Section: Resultsmentioning

confidence: 99%

A simple protein extraction method for proteomic analysis of mahogany (Swietenia macrophylla) embryos

et al. 2017

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In any proteomic studies, protein extraction and sample preparation are the most crucial steps for obtaining optimal results. This is to ensure extracted proteins are not only high in yield but also clean from contaminants that could affect downstream proteomic applications such as two dimensional gel electrophoresis (2-DE) and mass spectrometry. Tissues from plants and trees such as Swietenia macrophylla are often rich in non-protein contaminating substances, which could interfere in the proteomic applications. S. macrophylla or also known as the mahogany is one of the most valuable tree species in the world. Studies on proteins for this tree as well as its seeds are very limited. We have extracted proteins from S. macrophylla seeds (specifically embryo tissues) using three different methods, each having different lysis buffer recipes. Furthermore, another set of samples were precipitated using trichloroacetic acid/acetone prior to the three extraction methods to further purify the protein samples. The results from 2-DE analysis showed approximately 240 protein spots were detected from the successful protocol using a lysis buffer of 9 M urea, 4% CHAPS, 0.5% triton X-100 and 100 mM DTT without TCA/acetone precipitation. This study highlights the aspects of sample preparation for S. macrophylla embryos, focusing on the total protein extraction and resolution in SDS-PAGE as well as 2-DE. Furthermore, this is the very first report of the proteome 2DE profile from S. macrophylla embryo.

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“…However, little effort have been done on olive proteomic, probably because of the difficulty on sample handling, due to the presence of many phenolic compounds (Garcia et al 2000, Wang et al, 2003, Wang et al, 2004, Wang et al, 2010.…”

Section: Proteomic Changes In Developing Olive Inflorescencementioning

confidence: 99%

“…A modified protocol based on those suggested by Garcia et al (2000) Süle et al (2004), and Wang et al (2003and Wang et al ( , 2004 for olive was followed for protein extraction and purification. Proteins were extracted using a 10fold volume/weight buffer (50 mM sodium borate, 50 mM ascorbic acid pH 9.0, 1% β-mercaptoethanol, 1% soluble PVP, 1% insoluble PVP, 10 mM PMSF) under continuous vortexing for 60 min at 4 o C. The homogenate was centrifuged at 4 o C for 30 min at 35,000 g and the pellet was discarded.…”

Section: Proteomic Analysesmentioning

confidence: 99%

Nutritional and Proteomic Profiles in Developing Olive Inflorescence

Kitsaki¹,

Maragos²,

Bouranis³

2012

Advances in Selected Plant Physiology Aspects

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Removal of lipid contaminants by organic solvents from oilseed protein extract prior to electrophoresis

Cited by 39 publications

References 10 publications

Optimizing protein extraction from plant tissues for enhanced proteomics analysis

Optimizing protein extraction from plant tissues for enhanced proteomics analysis

A simple protein extraction method for proteomic analysis of mahogany (Swietenia macrophylla) embryos

Nutritional and Proteomic Profiles in Developing Olive Inflorescence

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