| 1. |
- Qi, Xiaoying, et al.
(author)
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High Throughput, Absolute Determination of the Content of a Selected Protein at Tissue Levels Using Quantitative Dot Blot Analysis (QDB)
- 2018
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In: Journal of Visualized Experiments. - : MyJove Corporation. - 1940-087X. ; :138
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Journal article (peer-reviewed)abstract
- Lacking a convenient, quantitative, high throughput immunoblot method for absolute determination of the content of a specific protein at cellular and tissue level significantly hampers the progress in proteomic research. Results derived from currently available immunoblot techniques are also relative, preventing any efforts to combine independent studies with a large-scale analysis of protein samples. In this study, we demonstrate the process of quantitative dot blot analysis (QDB) to achieve absolute quantification in a high throughput format. Using a commercially available protein standard, we are able to determine the absolute content of capping actin protein, gelsolin-like (CAPG) in protein samples prepared from three different mouse tissues (kidney, spleen, and prostate) together with a detailed explanation of the experimental details. We propose the QDB analysis as a convenient, quantitative, high throughput immunoblot method of absolute quantification of individual proteins at the cellular and tissue level. This method will substantially aid biomarker validation and pathway verification in various areas of biological and biomedical research.
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| 2. |
- Zhu, Yanping, et al.
(author)
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Identification of prothymosin alpha (PTMA) as a biomarker for esophageal squamous cell carcinoma (ESCC) by label-free quantitative proteomics and Quantitative Dot Blot (QDB)
- 2019
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In: Clinical Proteomics. - : BMC. - 1542-6416 .- 1559-0275. ; 16
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Journal article (peer-reviewed)abstract
- Background: Esophageal cancer (EC) is one of the malignant tumors with a poor prognosis. The early stage of EC is asymptomatic, so identification of cancer biomarkers is important for early detection and clinical practice.Methods: In this study, we compared the protein expression profiles in esophageal squamous cell carcinoma (ESCC) tissues and adjacent normal esophageal tissues from five patients through high-resolution label-free mass spectrometry. Through bioinformatics analysis, we found the differentially expressed proteins of ESCC. To perform the rapid identification of biomarkers, we adopted a high-throughput protein identification technique of Quantitative Dot Blot (QDB). Meanwhile, the QDB results were verified by classical immunohistochemistry.Results: In total 2297 proteins were identified, out of which 308 proteins were differentially expressed between ESCC tissues and normal tissues. By bioinformatics analysis, the four up-regulated proteins (PTMA, PAK2, PPP1CA, HMGB2) and the five down-regulated proteins (Caveolin, Integrin beta-1, Collagen alpha-2(VI), Leiomodin-1 and Vinculin) were selected and validated in ESCC by Western Blot. Furthermore, we performed the QDB and IHC analysis in 64 patients and 117 patients, respectively. The PTMA expression was up-regulated gradually along the progression of ESCC, and the PTMA expression ratio between tumor and adjacent normal tissue was significantly increased along with the progression. Therefore, we suggest that PTMA might be a potential candidate biomarker for ESCC.Conclusion: In this study, label-free quantitative proteomics combined with QDB revealed that PTMA expression was up-regulated in ESCC tissues, and PTMA might be a potential candidate for ESCC. Since Western Blot cannot achieve rapid and high-throughput screening of mass spectrometry results, the emergence of QDB meets this demand and provides an effective method for the identification of biomarkers.
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| 3. |
- Tian, Geng, et al.
(author)
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Quantitative dot blot analysis (QDB), a versatile high throughput immunoblot method
- 2017
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In: Oncotarget. - : Impact Journals, LLC. - 1949-2553. ; 8:35, s. 58553-58562
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Journal article (peer-reviewed)abstract
- Lacking access to an affordable method of high throughput immunoblot analysis for daily use remains a big challenge for scientists worldwide. We proposed here Quantitative Dot Blot analysis (QDB) to meet this demand. With the defined linear range, QDB analysis fundamentally transforms traditional immunoblot method into a true quantitative assay. Its convenience in analyzing large number of samples also enables bench scientists to examine protein expression levels from multiple parameters. In addition, the small amount of sample lysates needed for analysis means significant saving in research sources and efforts. This method was evaluated at both cellular and tissue levels with unexpected observations otherwise would be hard to achieve using conventional immunoblot methods like Western blot analysis. Using QDB technique, we were able to observed an age-dependent significant alteration of CAPG protein expression level in TRAMP mice. We believe that the adoption of QDB analysis would have immediate impact on biological and biomedical research to provide much needed high-throughput information at protein level in this "Big Data" era.
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| 4. |
- Moore, R. G., et al.
(author)
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Manifestations of broken symmetry : The surface phases of Ca2-xSrxRuO4
- 2008
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In: Physical Review Letters. - 0031-9007 .- 1079-7114. ; 100:6, s. 066102-
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Journal article (peer-reviewed)abstract
- The surface structural phases of Ca2-xSrxRuO4 are investigated using quantitative low energy electron diffraction. The broken symmetry at the surface enhances the structural instability against the RuO6 rotational distortion while diminishing the instability against the RuO6 tilt distortion occurring within the bulk crystal. As a result, suppressed structural and electronic surface phase transition temperatures are observed, including the appearance of an inherent Mott metal-to-insulator transition for x=0.1 and possible modifications of the surface quantum critical point near x(c)similar to 0.5.
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| 5. |
- Nascimento, V. B., et al.
(author)
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Procedure for LEED I-V structural analysis of metal oxide surfaces : Ca1.5Sr0.5RuO4(001)
- 2007
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In: Physical Review B. Condensed Matter and Materials Physics. - 1098-0121 .- 1550-235X. ; 75:3, s. 035408-
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Journal article (peer-reviewed)abstract
- Transition metal oxides (TMOs) are famous for the intimate coupling between the lattice, electrons, and spin, creating exotic functionality. Creating a surface, breaking the symmetry, should result in lattice distortions that due to the close coupling could create different "surface phases." Historically it has been very difficult to use low energy electron diffraction I-V to quantitatively determine the surface structure of TMOs. A signature of this problem is the large values commonly reported in the literature of the Pendry reliability factor (R-P), which is used to quantify the agreement between experimental data and calculated diffraction. In this paper we describe a consistent procedure for determining the phase shifts using an optimized muffin-tin potential approach combined with an energy-dependent real and imaginary inner potential. This procedure is used to determine the surface structure of the layered TMO Ca1.5Sr0.5RuO4. An acceptable Pendry reliability factor is achieved (R-P=0.28).
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