using the equation A = log(UT) convert: a, 83% transmittance to absorbance b. 0.
ID: 985809 • Letter: U
Question
using the equation A = log(UT) convert: a, 83% transmittance to absorbance b. 0.22 absorbance to transmittance. 1, 2. Concentration estimation: a. Using the graph in Figure 2 (standard plot), estimate the concentration of a sample with an absorbance of 0.2 b. Would the absorbance reading of the previous question be within the range of the spectrophotometer? 3. In what order do you use the knobs on the spectrophotometer during calibration? 4. A wavelength of 595 nm will be used for this assay. What is the expected observed color of the samples? 5. What is in your blank (reference cuvette) for this assay? 6. Serial dilution. See pre-laboratory procedure (below)Explanation / Answer
1)A = log (1/T)
83% transmittance to absorbance A = log(1/0.83) = 0.080
0.22 absorbance to transmittance .22 = log(1/t) = 0.60 = 60% transmittance
2)a) its around 15mg/ml
,b)yes because it is on the line which pass through origin and linearly dependent on concentration
3) to adjust the various parameters like the wavelenth where the absorbanse needs to be taken and to set measuure the reading at blank w need to adjust the knobs accordingly
4) blue color due to the dyes is converted to blue ater interaction with proteins
5) only dye and buffer but no protein is there
6)Prepare a series of protein standards diluted with 0.15 M NaCl to final concentrations of 0 (blank = NaCl only), 250, 500, 750 and 1500 µg/mL. Also prepare serial dilutions of the unknown sample to be measured.
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