Comparison of sample preparation methods suitable for poplar leaf proteomics

doi:10.3969/j.issn.1000-2006.201706063

ZHEN Yan, ZHENG Xiuhua, SHI Jisen

JOURNAL OF NANJING FORESTRY UNIVERSITY ›› 2018, Vol. 42 ›› Issue (02) : 33-39.

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JOURNAL OF NANJING FORESTRY UNIVERSITY ›› 2018, Vol. 42 ›› Issue (02) : 33-39. DOI: 10.3969/j.issn.1000-2006.201706063

Comparison of sample preparation methods suitable for poplar leaf proteomics

ZHEN Yan, ZHENG Xiuhua, SHI Jisen^*

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Abstract

【Objective】In order to improve the detection of low abundance proteins in plant leaves, the effective extraction methods were established in poplar leaves for proteomics analysis.【Method】 Trichloroacetic acid(TCA)-acetone coupled with polyethylene glycol(PEG)fractionation and sodium dodecyl sulfate(SDS)-phenol coupled with PEG fractionation were evaluated, combined with SDS-polyacrylamide gel electrophoresis(SDS-PAGE)and two-dimensional electrophoresis(2-DE).【Result】The SDS-PAGE results showed that TCA-acetone coupled with PEG fractionation is superior to SDS-phenol coupled with the PEG fractionation method. Further, proteins with PEG fractionation and no PEG fractionation coupled with TCA-acetone were compared by using 2-DE, and the gel maps displayed that PEG fractionation is a good way to deplete the Rubiscos and improve the detection of low abundant proteins. The PEG fractionation method can detect nearly 90 more spots than nonfractionation can. 【Conclusion】LTQ-Orbitrap XL was used to identify several proteins from the PEG fractionation coupled with TCA-acetone. The TCA-acetone coupled with PEG fractionation is an effective approach for the detection of 2-DE analysis and mass spectroscopy(MS)identification of lower abundance proteins.

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ZHEN Yan, ZHENG Xiuhua, SHI Jisen. Comparison of sample preparation methods suitable for poplar leaf proteomics[J]. JOURNAL OF NANJING FORESTRY UNIVERSITY. 2018, 42(02): 33-39 https://doi.org/10.3969/j.issn.1000-2006.201706063

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