User:Catherine Koenigsknecht/Notebook/Experimental Biological Chemistry/2011/10/18: Difference between revisions
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==Objective== | ==Objective== | ||
Make a labeled cysteine fluorophore. | |||
==Description== | ==Description== | ||
# | # Measure out 5mg of 5-(Iodoacetamido)fluorescein and put in a 50mL Falcon tube. | ||
# Add 19.6μL of 118mg/mL cysteine to the 50mL Falcon tube. | |||
# Add 35mL of 50mM Tris, pH 7.5. | |||
# Add 100μL of 2.25M DTT after about two hours of the reaction occurring at room temperature. | |||
==Data== | ==Data== | ||
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==Notes== | ==Notes== | ||
#Abs at 490nm = 1.144AU for 20x dilute | |||
*A=bcε, b=1cm, ε=42000M-1cm-1 | |||
*c=27.24μM for 20x dilute | |||
#actual concentration is 544.76μM | |||
*concentration was supposed to be 277.231μM (.005g/(515.3g/mol)/(.035L)*106) | |||
#Concentration of cysteine = ((118g/L)*(1.96*10-5L))/(121.16 g/mol)/(.035L)*106 = 545.395μM | |||
Revision as of 09:41, 26 December 2011
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Entry titleObjectiveMake a labeled cysteine fluorophore. Description
Data
Notes
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