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EV WP Or bcelynlirmes LABORATORY REPORT Name: Date: EXPERIMENTAL NOTES COMPLETE THE FOLLOWING TABLE TABLE 9.1 PROTEIN OR SUBSTANCE INDICATE THE WAVELENGTH(S) IN WHICH THE ABSORBANCE OF THE PROTEIN OR SUBSTANCE WILL BE MONITORED GY0 nm 280 280 Blue Dextran Cytochrome e 400 nm AND Ovalbumin hm Vitamin B12 3 60 nm STANDARDS DATA FROM GEL FILTRATION COLUMN V for Blue Dextran = 17.3mL V for vitamin B12 = 40.3mL TABLE 9.2 V. MOLECULAR WEIGHT PROTEIN av -0.068 53,000 dalfom 34,620 dlfans 13, 700 do ltans G, 311 daltons 19.3mL GLUTAMATE DEHYDROGENASE 22.3mL PEPSIN 28.6mL RNASE 33.7mL APROTININ - 103 - PARTI 1.2 Abs 280n m 0.8 Abs 360nm 1- Abs 400nm 0.6 Vo Abs 640nm 0.4 Vt 0.46 0.2 6,28 . 10 20 V 30 40 50 60 Elution Volume, ml 46 FIGURE 9.3 Elution profile on a Sephacryl S200 column CALCULATING THE MOLECULAR WEIGHT OF AN UNKNOWN Using the information provided by such a graph, one can calculate the molecular weight of an unknown component in a sample. This is done by first running a sample containing a set of known standards along with a protein which will run in the void volume and define the V of the column. The V. of the column can be either calculated, or defined by a very small molecule as described above. Using these values, one can then calculate the distribution coefficient, or K formula for calculation of K is (K) of each known standard. The average av av V.-V. V-V, Ky av Absorbance

EV WP Or bcelynlirmes LABORATORY REPORT Name: Date: EXPERIMENTAL NOTES COMPLETE THE FOLLOWING TABLE TABLE 9.1 PROTEIN OR SUBSTANCE INDICATE THE WAVELENGTH(S) IN WHICH THE ABSORBANCE OF THE PROTEIN OR SUBSTANCE WILL BE MONITORED GY0 nm 280 280 Blue Dextran Cytochrome e 400 nm AND Ovalbumin hm Vitamin B12 3 60 nm STANDARDS DATA FROM GEL FILTRATION COLUMN V for Blue Dextran = 17.3mL V for vitamin B12 = 40.3mL TABLE 9.2 V. MOLECULAR WEIGHT PROTEIN av -0.068 53,000 dalfom 34,620 dlfans 13, 700 do ltans G, 311 daltons 19.3mL GLUTAMATE DEHYDROGENASE 22.3mL PEPSIN 28.6mL RNASE 33.7mL APROTININ - 103 - PARTI 1.2 Abs 280n m 0.8 Abs 360nm 1- Abs 400nm 0.6 Vo Abs 640nm 0.4 Vt 0.46 0.2 6,28 . 10 20 V 30 40 50 60 Elution Volume, ml 46 FIGURE 9.3 Elution profile on a Sephacryl S200 column CALCULATING THE MOLECULAR WEIGHT OF AN UNKNOWN Using the information provided by such a graph, one can calculate the molecular weight of an unknown component in a sample. This is done by first running a sample containing a set of known standards along with a protein which will run in the void volume and define the V of the column. The V. of the column can be either calculated, or defined by a very small molecule as described above. Using these values, one can then calculate the distribution coefficient, or K formula for calculation of K is (K) of each known standard. The average av av V.-V. V-V, Ky av Absorbance

Fundamentals Of Analytical Chemistry
Fundamentals Of Analytical Chemistry
9th Edition
ISBN:9781285640686
Author:Skoog
Publisher:Skoog
Chapter25: Instruments For Optical Spectrometry
Section: Chapter Questions
Problem 25.14QAP
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When trying to calculate the Kav for glutamate dehydrogenase I keep getting a negative value using the given values along with those in the graph but I fear that that's not correct

EV WP Or bcelynlirmes LABORATORY REPORT Name: Date: EXPERIMENTAL NOTES COMPLETE THE FOLLOWING TABLE TABLE 9.1 PROTEIN OR SUBSTANCE INDICATE THE WAVELENGTH(S) IN WHICH THE ABSORBANCE OF THE PROTEIN OR SUBSTANCE WILL BE MONITORED GY0 nm 280 280 Blue Dextran Cytochrome e 400 nm AND Ovalbumin hm Vitamin B12 3 60 nm STANDARDS DATA FROM GEL FILTRATION COLUMN V for Blue Dextran = 17.3mL V for vitamin B12 = 40.3mL TABLE 9.2 V. MOLECULAR WEIGHT PROTEIN av -0.068 53,000 dalfom 34,620 dlfans 13, 700 do ltans G, 311 daltons 19.3mL GLUTAMATE DEHYDROGENASE 22.3mL PEPSIN 28.6mL RNASE 33.7mL APROTININ - 103 - PARTI
Transcribed Image Text:EV WP Or bcelynlirmes LABORATORY REPORT Name: Date: EXPERIMENTAL NOTES COMPLETE THE FOLLOWING TABLE TABLE 9.1 PROTEIN OR SUBSTANCE INDICATE THE WAVELENGTH(S) IN WHICH THE ABSORBANCE OF THE PROTEIN OR SUBSTANCE WILL BE MONITORED GY0 nm 280 280 Blue Dextran Cytochrome e 400 nm AND Ovalbumin hm Vitamin B12 3 60 nm STANDARDS DATA FROM GEL FILTRATION COLUMN V for Blue Dextran = 17.3mL V for vitamin B12 = 40.3mL TABLE 9.2 V. MOLECULAR WEIGHT PROTEIN av -0.068 53,000 dalfom 34,620 dlfans 13, 700 do ltans G, 311 daltons 19.3mL GLUTAMATE DEHYDROGENASE 22.3mL PEPSIN 28.6mL RNASE 33.7mL APROTININ - 103 - PARTI
1.2 Abs 280n m 0.8 Abs 360nm 1- Abs 400nm 0.6 Vo Abs 640nm 0.4 Vt 0.46 0.2 6,28 . 10 20 V 30 40 50 60 Elution Volume, ml 46 FIGURE 9.3 Elution profile on a Sephacryl S200 column CALCULATING THE MOLECULAR WEIGHT OF AN UNKNOWN Using the information provided by such a graph, one can calculate the molecular weight of an unknown component in a sample. This is done by first running a sample containing a set of known standards along with a protein which will run in the void volume and define the V of the column. The V. of the column can be either calculated, or defined by a very small molecule as described above. Using these values, one can then calculate the distribution coefficient, or K formula for calculation of K is (K) of each known standard. The average av av V.-V. V-V, Ky av Absorbance
Transcribed Image Text:1.2 Abs 280n m 0.8 Abs 360nm 1- Abs 400nm 0.6 Vo Abs 640nm 0.4 Vt 0.46 0.2 6,28 . 10 20 V 30 40 50 60 Elution Volume, ml 46 FIGURE 9.3 Elution profile on a Sephacryl S200 column CALCULATING THE MOLECULAR WEIGHT OF AN UNKNOWN Using the information provided by such a graph, one can calculate the molecular weight of an unknown component in a sample. This is done by first running a sample containing a set of known standards along with a protein which will run in the void volume and define the V of the column. The V. of the column can be either calculated, or defined by a very small molecule as described above. Using these values, one can then calculate the distribution coefficient, or K formula for calculation of K is (K) of each known standard. The average av av V.-V. V-V, Ky av Absorbance
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