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What is the concentration of carnosine in your cell lysate?

What is the concentration of carnosine in your cell lysate?

Biochemistry
Biochemistry
9th Edition
ISBN:9781319114671
Author:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Publisher:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Chapter1: Biochemistry: An Evolving Science
Section: Chapter Questions
Problem 1P
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1. This question is adapted from Harris 19-D. Carnosine is a dipeptide whose antioxidant properties protect cells from free radicals. Carnosine was determined by derivatization with naphthalene-2,3-dicarboxyaldehyde and cyanide followed by fluorescence detection using excitation at 445 nm and emission at 490. Quantification was by standard addition. To four aliquots of 20-ul cell lysate were added volumes of 100µM carnosine standard to generate final concentrations of 0, 1.0, 2.5, and 5.0 µM added carnosine. Solutions were diluted to 70µl before addition of 15µl of 5 mM naphthalene-2,3-dicarboxyaldehyde and 15µl of 10 mM NaCN. H H2N. OH Carnosine HN. Naphthalene-2,3- dicarboxyaldehyde Cyanide OH Fluorescent product HN. Fluorescence intensity Concentration (uM) of added carnosine in final 100 µL 0.0 0.465 1.0 0.698 2.5 1.029 5.0 1.651
Transcribed Image Text:1. This question is adapted from Harris 19-D. Carnosine is a dipeptide whose antioxidant properties protect cells from free radicals. Carnosine was determined by derivatization with naphthalene-2,3-dicarboxyaldehyde and cyanide followed by fluorescence detection using excitation at 445 nm and emission at 490. Quantification was by standard addition. To four aliquots of 20-ul cell lysate were added volumes of 100µM carnosine standard to generate final concentrations of 0, 1.0, 2.5, and 5.0 µM added carnosine. Solutions were diluted to 70µl before addition of 15µl of 5 mM naphthalene-2,3-dicarboxyaldehyde and 15µl of 10 mM NaCN. H H2N. OH Carnosine HN. Naphthalene-2,3- dicarboxyaldehyde Cyanide OH Fluorescent product HN. Fluorescence intensity Concentration (uM) of added carnosine in final 100 µL 0.0 0.465 1.0 0.698 2.5 1.029 5.0 1.651
c) What is the concentration of carnosine in your cell lysate? If you use excel at any point, please include a copy of your workings.
Transcribed Image Text:c) What is the concentration of carnosine in your cell lysate? If you use excel at any point, please include a copy of your workings.
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