I have estimated proline in sugar beet leaves and have observed the absorbance at 520 nm. Now I want to make a standard curve for further estimation and calculation of proline. Can anyone help me how to prepare standard curve for the same.
Read the following papers:
Lotfi N, Vahdati K, Kholdebarin B and Amiri R (2009) Soluble sugars and proline accumulation play a role as effective indices for drought tolerant screening in Persian walnut (Juglans regia L.) during germination. Fruits. 65(2):1-14.
Aslani Aslamarz AA, Vahdati K, Hasani D, Rahemi M, Leslie CA (2011) Cold hardiness and its relationship with proline content in Persian walnut. European Journal of Horticultural Sciences. 76 (3): 84–90.
Jariteh M, Ebrahimzadeh H, Niknam V, Mirmasoumi M and Vahdati K(2015) Developmental changes of protein, proline and some antioxidant enzymes activities in somatic and zygotic embryos of Persian walnut (Juglans regiaL.). Plant, Cell, Tissue and Organ Culture. 122:101–115.
Lotfi N, Vahdati K, Kholdebarin B and Amiri R (2010) Drought-induced accumulation of sugars and proline in radicle and plumule of tolerant walnut varieties during germination phase. Acta Horticulturae. 861:289-296.
Sir there is mention of the proline estimation by Bates method which I knew but there is no mention of how the standard curve is made. It says that L proline is used for standard estimation. I wanted to know how much known concentration of stock of L proline will be made. How we will calculate the amount and at what different known concentration of L proline is standard curve is made.
Thank you Sir. I know about Bates Method. I was talking about the concentration and all
If I am not too late to reply Varucha Misra
Preparation of standard curve:
L-proline standard solutions 0.0, 0.20, 0.40, 0.60, 0.80, 1.00 mL (0, 5, 10, 15, 20, 25 μg L-proline) were added to tubes with stoppers respectively and were all diluted with water to 1 ml
Follow Bates method (1973) to quantify @520nm
Thank you. Will definitely try this also. Though we tried by other concentration.
My intercept value was more than OD value..So, many proline concentration was negative...What does it mean?
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