Quantification of Total and Soluble Inorganic Phosphate
植物中总无机磷酸盐和可溶性无机磷酸盐的定量分析
发布: 2013年09月05日第3卷第17期 DOI: 10.21769/BioProtoc.890 浏览次数: 15766
评审: Tie Liu
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参见作者原研究论文
The authors used this protocol in:
Jul 2010
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Abstract
A simple, rapid, and sensitive colorimetric microassay for inorganic phosphate (Pi) relies upon the absorption at 660 nm of a molybdenum blue complex that forms upon reduction of an ammonium molybdate-Pi complex in acid. The method for determination of total Pi uses plant tissues that have been ashed at 500 °C, whereas quantification of soluble Pi is performed with tissues extracted under mild acid conditions (which preserves acid-labile phosphate ester bonds).
Materials and Reagents
- Plant tissues
- 17.5 M Glacial acetic acid
- 12 M Concentrated HCl
- 16 M Concentrated HNO3
- Sodium phosphate, monobasic (NaH2PO4) (Bioshop, catalog number: SPM400 )
- Ascorbic acid (Bioshop, catalog number: AS0704 )
- Ammonium molybdate (Bioshop, catalog number: AMN333 )
- Zinc acetate (Sigma-Aldrich, catalog number: Z0625 )
- Quartz crucibles (Thermo Fisher Scientific, catalog number: 08-072 series)
- Acid extraction solution (see Recipes)
- Pi stock (for standard curve) (see Recipes)
- Pi assay reagent (see Recipes)
Equipment
- Crucible
- Drying oven
- Repeat pipetor
- Isotemp Muffle Furnace (Thermo Fisher Scientific, model: 10-650-14 )
- Microcentrifuge
- A computer supported microplate spectrophotometer (e.g., Spectromax Plus, Molecular Devices, Sunnyvale)
Procedure
文章信息
版权信息
© 2013 The Authors; exclusive licensee Bio-protocol LLC.
如何引用
Readers should cite both the Bio-protocol article and the original research article where this protocol was used:
- Knowles, V. and Plaxton, W. (2013). Quantification of Total and Soluble Inorganic Phosphate. Bio-protocol 3(17): e890. DOI: 10.21769/BioProtoc.890.
- Hurley, B. A., Tran, H. T., Marty, N. J., Park, J., Snedden, W. A., Mullen, R. T. and Plaxton, W. C. (2010). The dual-targeted purple acid phosphatase isozyme AtPAP26 is essential for efficient acclimation of Arabidopsis to nutritional phosphate deprivation. Plant Physiol 153(3): 1112-1122.
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分类
植物科学 > 植物生物化学 > 其它化合物
生物化学 > 其它化合物 > 离子 > 磷酸根
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