Quantitative Analysis of Iron in Protein Bands Existing in Polyacrylamide Gel Matrix with Synchrotron Radiation

LIU Ying-Bin; GAO Yu-Xi; CHEN Chun-Ying; LI Bai; YU Hong-Wei; WANG Jiang-Xue; HE Wei; HUANG Yu-Ying; CHAI Zhi-Fang

Chinese Physics C> 2005, Vol. 29> Issue(S1) : 61-64

Quantitative Analysis of Iron in Protein Bands Existing in Polyacrylamide Gel Matrix with Synchrotron Radiation

Institute of High Energy Physics,CAS,Beijing 100049,China2 Second Affiliated Hospital of Zhejiang University,Hangzhou 310009,China

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Abstract：
A procedure has been proposed for quantification of Fe in protein bands in polyacrylamide gel matrix with synchrotron radiation X-ray fluorescence(SRXRF)analysis. Human hemoglobin was separated with isoelectric focusing (IEF), and Fe amounts in protein bands were measured with SRXRF. Calibration standards were prepared by electrophoresis of bovine serum albumin containing known amounts of Fe with stacking gel. The calibration curves can be obtained in a range of 0—5.1ng Fe and linear relationship between dosage of metals and fluorescent intensity can be observed (R²=0.9897). The method provides detection limit of 0.63ng Fe,and the recovery of Fe determination is about 106%. The hyphenated technique of SRXRF and IEF has advantages of quantifying trace elements in proteins and identifying the proteins simultaneously. It can be employed for speciation analysis of trace elements in biological samples.

References

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LIU Ying-Bin, GAO Yu-Xi, CHEN Chun-Ying, LI Bai, YU Hong-Wei, WANG Jiang-Xue, HE Wei, HUANG Yu-Ying and CHAI Zhi-Fang. Quantitative Analysis of Iron in Protein Bands Existing in Polyacrylamide Gel Matrix with Synchrotron Radiation[J]. Chinese Physics C, 2005, 29(S1): 61-64.

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Received: 2005-10-28
Revised: 1900-01-01

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通讯作者: 陈斌, bchen63@163.com

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沈阳化工大学材料科学与工程学院沈阳 110142

Quantitative Analysis of Iron in Protein Bands Existing in Polyacrylamide Gel Matrix with Synchrotron Radiation

Corresponding author: GAO Yu-Xi,

Institute of High Energy Physics,CAS,Beijing 100049,China2 Second Affiliated Hospital of Zhejiang University,Hangzhou 310009,China

Received Date: 2005-10-28

Accepted Date: 1900-01-01

Available Online: 2005-01-02

Abstract: A procedure has been proposed for quantification of Fe in protein bands in polyacrylamide gel matrix with synchrotron radiation X-ray fluorescence(SRXRF)analysis. Human hemoglobin was separated with isoelectric focusing (IEF), and Fe amounts in protein bands were measured with SRXRF. Calibration standards were prepared by electrophoresis of bovine serum albumin containing known amounts of Fe with stacking gel. The calibration curves can be obtained in a range of 0—5.1ng Fe and linear relationship between dosage of metals and fluorescent intensity can be observed (R²=0.9897). The method provides detection limit of 0.63ng Fe,and the recovery of Fe determination is about 106%. The hyphenated technique of SRXRF and IEF has advantages of quantifying trace elements in proteins and identifying the proteins simultaneously. It can be employed for speciation analysis of trace elements in biological samples.

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Quantitative Analysis of Iron in Protein Bands Existing in Polyacrylamide Gel Matrix with Synchrotron Radiation

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