UV-1800_user_guide_Ch12

UV-1800_user_guide_Ch12 - 12 12 Bio-method 12 1 1 Chapter...

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UV-1800 SERIES 12-1 12 12 1 1 1 1 1 1 1 1 1 12 12 12 12 12 12 12 12 Bio-method Chapter 12 Bio-method The Bio-method mode allows you to obtain the DNA and protein concentrations with various quantitation methods. CONTENTS 12.1 Bio-method Selection . ..................................................................................................................... 12-2 12.2 DNA Quantitation. ............................................................................................................................ 12-4 12.3 Lowry Method, BCA Method, CBB Method, and Biuret Method. ................................................... 12-14 12.4 UV Absorption Method . ................................................................................................................. 12-16
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UV-1800 SERIES 12-2 1212 1212 12 12 12 12 12 12 12 12 12 12 12 12 12 12 12 12 12.1 12.1 Bio-method Selection When you select [7.Bio-Method] in the [Mode menu] screen ( Fig. 2.1 ), the screen for selecting a quantitation method appears ( Fig. 12.1 ). Select the item number for a desired method with numeric keys while the selection screen ( Fig. 12.1 ) is displayed. Fig. 12.1 Bio-Method Selection screen No. Key Operation Display Description 1 [DNA Quantitation] [DNA Quantitation] performs measurement at specified two to three fixed wavelengths and obtains the concentration of DNA and proteins, and absorbance ratio, based on the measured absorbances. You can choose any wavelength for measurements, including the most commonly used wavelengths (260 nm/ 230 nm or 260 nm/ 280 nm). You can add measurements at a wavelength of 320 nm for back ground correction. You can also set the desired parameters for calculating the DNA and protein concentrations in samples. ^ "12.2 DNA Quantitation" 2 [Lowry Method] This method offers excellent sensitivity and reproducibility. Based on reactions to tyrosine and tryptophan, this method utilizes different absorbance values depending on the type of protein measured, even if the amount of protein is the same. ^ "12.3 Lowry Method, BCA Method, CBB Method, and Biuret Method" 1 2 3 4 5 6
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12.1 Bio-method Selection UV-1800 SERIES 12-3 12 12 1 1 1 1 1 1 1 1 1 12 12 12 12 12 12 12 For details on the quantitation methods described above, refer to the following references. 1. Warberg, O., and Christian, W. (1942) Biochem. Z.310, 384-421. 2. Vernon F. Kalb, Jr., and Robert W. Bernlohr (1977) Anal. Biochem. 82, 362-371. 3. Gornall, A.G., Bradawill, C. J. & David, M.M. (1949) J. Biol. Chem. 177, 751-766. 4. Lowry, O. H., Rosebrough, N. J., Farr, A. I. & Randall, R. J. (1951) J. Biol. Chem. 193, 265-275. 5. Bradford, M. M. (1976) Anal. Biochem. 72, 248-254. 6. Smith, P. K., Krohn, R. I., Hermanson, G. T., Mallia, A. K., Gartner, F. H., Provenzano, M. D., Fujimoto, E. K., Goeke, N. M., Olson, B. J. & Klenk, D. C. (1985) Anal. Biochem. 150, 76-85. 3 [BCA Method] This method measures the absorbance at a wavelength of 562 nm using a reagent called Bicinchoninic Acid. You must take into consideration that the absorbance is known to increase with time when this method is used. ^ "12.3 Lowry Method, BCA Method, CBB Method, and Biuret Method" 4 [CBB Method (Bradford Method)] This method measures the absorbance at a wavelength of 595 nm using a reagent called Coomassie Brilliant Blue G-250. ^ "12.3 Lowry Method, BCA Method, CBB Method, and Biuret Method" 5 [Biuret Method] This method is based on the unique peptide linkage of protein as well as the Biuret reaction which involves the formation of a colored compound with copper reagent. This method offers a fast and easy determination of protein concentration. The drawback is its low sensitivity. In this method, absorbance is normally measured at a wavelength of 540 to 560 nm.
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UV-1800_user_guide_Ch12 - 12 12 Bio-method 12 1 1 Chapter...

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