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Analytical technique for label-free multi-protein detection based on Western blot and surface-enhanced Raman scattering.
Anal Chem. 2008 Apr 15; 80(8):2799-804.AC

Abstract

We have developed a new analytical procedure for label-free protein detection designated "Western SERS", consisting of protein electrophoresis, Western blot, colloidal silver staining, and surface-enhanced Raman scattering (SERS) detection. A novel method of silver staining for Western blot that uses a silver colloid, an excellent SERS-active substrate, is first proposed in the present study. During the process of silver staining, interactions between proteins and silver nanoparticles result in the emergence of SERS of proteins. In the present study, we use myoglobin (Mb) and bovine serum albumin (BSA) as model proteins. From different protein bands on a nitrocellulose (NC) membrane, we have observed surface-enhanced resonance Raman scattering (SERRS) spectra of Mb and SERS spectra of BSA. The proposed technique offers dual advantages of simplicity and high sensitivity. On one hand, after the colloidal silver staining, we can detect label-free multi-proteins directly on a NC membrane without digestion, extraction, and other pretreatments. On the other hand, the detection limit of the Western SERS is almost consistent with the detection limit of colloidal silver staining, and the SERRS detection limit of Mb is found to be 4 ng/band. This analytical method, which combines the technique of protein separation with SERS, may be a powerful protocol for label-free protein detection in proteomic research.

Authors+Show Affiliations

State Key Laboratory of Supramolecular Structure and Materials, Jilin University, Changchun 130012, PR China.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

18290672

Citation

Han, Xiao X., et al. "Analytical Technique for Label-free Multi-protein Detection Based On Western Blot and Surface-enhanced Raman Scattering." Analytical Chemistry, vol. 80, no. 8, 2008, pp. 2799-804.
Han XX, Jia HY, Wang YF, et al. Analytical technique for label-free multi-protein detection based on Western blot and surface-enhanced Raman scattering. Anal Chem. 2008;80(8):2799-804.
Han, X. X., Jia, H. Y., Wang, Y. F., Lu, Z. C., Wang, C. X., Xu, W. Q., Zhao, B., & Ozaki, Y. (2008). Analytical technique for label-free multi-protein detection based on Western blot and surface-enhanced Raman scattering. Analytical Chemistry, 80(8), 2799-804. https://doi.org/10.1021/ac702390u
Han XX, et al. Analytical Technique for Label-free Multi-protein Detection Based On Western Blot and Surface-enhanced Raman Scattering. Anal Chem. 2008 Apr 15;80(8):2799-804. PubMed PMID: 18290672.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Analytical technique for label-free multi-protein detection based on Western blot and surface-enhanced Raman scattering. AU - Han,Xiao X, AU - Jia,Hui Y, AU - Wang,Yan F, AU - Lu,Zhi C, AU - Wang,Chun X, AU - Xu,Wei Q, AU - Zhao,Bing, AU - Ozaki,Yukihiro, Y1 - 2008/02/22/ PY - 2008/2/23/pubmed PY - 2008/5/21/medline PY - 2008/2/23/entrez SP - 2799 EP - 804 JF - Analytical chemistry JO - Anal Chem VL - 80 IS - 8 N2 - We have developed a new analytical procedure for label-free protein detection designated "Western SERS", consisting of protein electrophoresis, Western blot, colloidal silver staining, and surface-enhanced Raman scattering (SERS) detection. A novel method of silver staining for Western blot that uses a silver colloid, an excellent SERS-active substrate, is first proposed in the present study. During the process of silver staining, interactions between proteins and silver nanoparticles result in the emergence of SERS of proteins. In the present study, we use myoglobin (Mb) and bovine serum albumin (BSA) as model proteins. From different protein bands on a nitrocellulose (NC) membrane, we have observed surface-enhanced resonance Raman scattering (SERRS) spectra of Mb and SERS spectra of BSA. The proposed technique offers dual advantages of simplicity and high sensitivity. On one hand, after the colloidal silver staining, we can detect label-free multi-proteins directly on a NC membrane without digestion, extraction, and other pretreatments. On the other hand, the detection limit of the Western SERS is almost consistent with the detection limit of colloidal silver staining, and the SERRS detection limit of Mb is found to be 4 ng/band. This analytical method, which combines the technique of protein separation with SERS, may be a powerful protocol for label-free protein detection in proteomic research. SN - 1520-6882 UR - https://www.unboundmedicine.com/medline/citation/18290672/Analytical_technique_for_label_free_multi_protein_detection_based_on_Western_blot_and_surface_enhanced_Raman_scattering_ L2 - https://doi.org/10.1021/ac702390u DB - PRIME DP - Unbound Medicine ER -