- #1
Sitix
- 2
- 0
Somebody please help me.
Practical:
A calibration graph should be prepared by taking 0.0 (zero, blank), 1.0, 2.0 and 3.0 ml aliquots of an aqueous solution containing both 1.67 mM D-glucose and 1.67 mM D-fructose. Add distilled water to maket it 3 ml. Add 1 ml DNS reagent. Incubate 5 min. Add 3 ml distilled water.Measure with spectrophotometer 540 nm
Do the same with inhibitors
Results:
Standard curve
ml glucose/fructose Absorbance
0,----- 0.0
1,----- 0.69
2,----- 1.44
3,----- 2.15
absorbance
ml sucrose No inhibitor + fructose +urea
0------------0.03------ 0.63------ 0.02
0.25-------- 1.08------ 0.97- ----- 0.43
0.5--------- 1.15------ 1.17- ----- 0.71
1.0--------- 2.29------ 1.71------- 1.02
1.5--------- 2.20------ 2.11------ 1.14
Questions
The standard curve drawn in terms of the molar concentration (in the 3 ml samples; not the volumes of standard mixture) of inverted sucrose (glucose + fructose) in the assay tubes.
From your results, determine:
1) the initial rates of reactions in units of mmole min-1 (ml diluted enzyme)-1
2) the Km and Vmax of the diluted enzyme, using the Lineweaver-Burk plot
3) determine the mode of inhibition for fructose and urea, and their Ki values using the Lineweaver-Burk plot
Please help me
I read around a lot but I just cannot start it
I wanted to use C1v1=C2V2 But I don't have the concentration
Please somebody at least tell me how to start it
Practical:
A calibration graph should be prepared by taking 0.0 (zero, blank), 1.0, 2.0 and 3.0 ml aliquots of an aqueous solution containing both 1.67 mM D-glucose and 1.67 mM D-fructose. Add distilled water to maket it 3 ml. Add 1 ml DNS reagent. Incubate 5 min. Add 3 ml distilled water.Measure with spectrophotometer 540 nm
Do the same with inhibitors
Results:
Standard curve
ml glucose/fructose Absorbance
0,----- 0.0
1,----- 0.69
2,----- 1.44
3,----- 2.15
absorbance
ml sucrose No inhibitor + fructose +urea
0------------0.03------ 0.63------ 0.02
0.25-------- 1.08------ 0.97- ----- 0.43
0.5--------- 1.15------ 1.17- ----- 0.71
1.0--------- 2.29------ 1.71------- 1.02
1.5--------- 2.20------ 2.11------ 1.14
Questions
The standard curve drawn in terms of the molar concentration (in the 3 ml samples; not the volumes of standard mixture) of inverted sucrose (glucose + fructose) in the assay tubes.
From your results, determine:
1) the initial rates of reactions in units of mmole min-1 (ml diluted enzyme)-1
2) the Km and Vmax of the diluted enzyme, using the Lineweaver-Burk plot
3) determine the mode of inhibition for fructose and urea, and their Ki values using the Lineweaver-Burk plot
Please help me
I read around a lot but I just cannot start it
I wanted to use C1v1=C2V2 But I don't have the concentration
Please somebody at least tell me how to start it