title: Comparative analyses of universal extraction buffers for assay of stress related biochemical and physiological parameters.
abstract: Comparative efficiency of three extraction solutions, including the universal sodium phosphate buffer (USPB), the Tris-HCl buffer (UTHB), and the specific buffers, were compared for assays of soluble protein, free proline, superoxide radical (O2∙-), hydrogen peroxide (H2O2), and the antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT), guaiacol peroxidase (POD), ascorbate peroxidase (APX), glutathione peroxidase (GPX), and glutathione reductase (GR) in Populus deltoide. Significant differences for protein extraction were detected via sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and two-dimensional electrophoresis (2-DE). Between the two universal extraction buffers, the USPB showed higher efficiency for extraction of soluble protein, CAT, GR, O2∙-, GPX, SOD, and free proline, while the UTHB had higher efficiency for extraction of APX, POD, and H2O2. When compared with the specific buffers, the USPB showed higher extraction efficiency for measurement of soluble protein, CAT, GR, and O2∙-, parallel extraction efficiency for GPX, SOD, free proline, and H2O2, and lower extraction efficiency for APX and POD, whereas the UTHB had higher extraction efficiency for measurement of POD and H2O2. Further comparisons proved that 100 mM USPB buffer showed the highest extraction efficiencies. These results indicated that USPB would be suitable and efficient for extraction of soluble protein, CAT, GR, GPX, SOD, H2O2, O2∙-, and free proline.
authors.affiliation: a Key Laboratory of Plant Germplasm Enhancement and Specialty Agriculture , Wuhan Botanical Garden, Chinese Academy of Sciences , Wuhan , Hubei , P. R. China.
authors.forename: Chunyu
authors.initials: C
authors.lastname: Han
authors.forename: Zhulong
authors.initials: Z
authors.lastname: Chan
authors.forename: Fan
authors.initials: F
authors.lastname: Yang
mesh_terms: D000975:Antioxidants / Q000737:chemistry / Q000302:isolation & purification*; D060387:Ascorbate Peroxidases / Q000737:chemistry / Q000302:isolation & purification; D002021:Buffers; D002374:Catalase / Q000737:chemistry / Q000302:isolation & purification; D005980:Glutathione Reductase / Q000737:chemistry* / Q000302:isolation & purification; D006861:Hydrogen Peroxide / Q000737:chemistry; D018384:Oxidative Stress; D009195:Peroxidase / Q000737:chemistry / Q000302:isolation & purification; D013482:Superoxide Dismutase / Q000737:chemistry / Q000302:isolation & purification
chemical_list: D000975:Antioxidants; D002021:Buffers; D006861:Hydrogen Peroxide; C121048:guaiacol peroxidase; D060387:Ascorbate Peroxidases; D002374:Catalase; D009195:Peroxidase; D013482:Superoxide Dismutase; D005980:Glutathione Reductase
keywords: antioxidant enzymes; extraction efficiency; free proline; reactive oxygen species; universal Tris-HCl buffer extraction solution; universal sodium phosphate buffer extraction solution