Proline accumulates in many plant species when exposed to environmental stresses such as water shortage, salinity, extreme temperatures, heavy metals, and high light intensity. Determination of proline content not only can monitor physiological status, but also is essential for plant breeding to screen plant varieties with improved salinity and drought tolerance.
With excellent experts in plant physiology and plant biochemistry, Lifeasible has developed a broad range of methods for proline content measurement, including:
- Acid-ninhydrin method. Proline in plants can react with acidic-ninhydrin to form a stable red-colored compound with a peak absorbance at 520 nm.
- Fluorescamine method. Fluorescamine can react with proline to yield a product with a strong absorption at 312 nm.
- Fluorescence competition method. Proline can competitively inhibit the fluorogenic reaction of primary amines with fludescamine. The degree of inhibition is proportional to the amount of proline, and can be measured at Ex/Em = 380/480 nm.
- Isatin paper assay. Isatin is a highly specific color reagent that can bind to proline and form a blue derivative -- pyrrole blue. The proline concentration can be reflected by the blue color intensity.
- High performance liquid chromatography (HPLC) method. 9-fluorenylmethyl-chloroformate (FMOC-Cl) can react with proline to produce stable and highly fluorescent derivatives. These derivatives can be separated by HPLC, and the fluorescence can be detected at Ex/Em = 260/315 nm.
- Optical method. The optical method is based on the measurement value angle (α) of the polarization plane of radiation at a certain wavelength λ. When increasing the proline concentration in solution, the angle of the polarization plane will increase. According to the standard curve with known proline concentration within c = (0.001 – 0.1) %, the proline content in the sample can be calculated.
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