lab4datacollection_1

lab4datacollection_1 - 3.5 ml Use the control tube to blank...

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Activity 2A: Standard Curve Tube # Absorbance Blank 0 1 0.03 0.15 2 0.05 0.37 3 0.08 0.61 4 0.1 1.08 5 0.13 1.25 6 0.15 1.5 The tubes for the standard curve were made by mixing known concentrations of L-cysteine with DTNB to produce TNB anion (yellow). Concentration of cysteine (mM) The mM of product thiocholine can be extrapolated from this curve because thiocholine has one sulfhydryl group just as cysteine and reacts similarly with DTNB to produce TNB anion. R-S represents L-cysteine or thiocholine clear 0 0.02 0.04 0.06 0 0.2 0.4 0.6 0.8 1 1.2 1.4 1.6 f(x) = 11.27x - 0.16 R² = 0.98 STAND L A412
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DTNB reacts with sulfhydryl containing compounds, represented here by R-S, to produce the yellow product TNB2- ( Pierce Biotechnology, Inc. 2004). clear yellow
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0.08 0.1 0.12 0.14 0.16 ARD CURVE L-CYSTEINE [mM]
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Sec# Group# Activity 2B Baseline Experiment Protocol Error: Please note that enzyme is 0.5 ug/ml not 5 ug/ml. Table 4.1 A 1 mg/ml stock is diluted 2000 fold for use in the assay. Reaction Tubes Control Experimental 0.1 M PB7 3.6 ml
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Unformatted text preview: 3.5 ml Use the control tube to blank the machine 7.5 mM ATCh Use micropipettes to add reagents 5 mM DTNB Total volume 4 ml 4 ml Abs. value expected AChE Activity: Baseline Experiment Reaction conditions Time (sec) A412 nm mM thiocholine Clock Time: 30 temp: 60 90 120 150 180 pH: 210 240 270 300 Group Members: Baseline Experiment Notes: 200 μ l 200 μ l 200 μ l 200 μ l 0.5 ug/ ml AChE (enzyme) 0 μ l 100 μ l Absorbance of Control tube at end of the reaction = Sec# AChE Activity: Baseline Experiment Activity 2B Class Data & Data Analysis Time (sec) Group 1 - A412 nm Group 2 - A412 nm Group 3 - A412 nm Group 4 - A412 nm 30 60 90 120 150 180 210 240 270 300 Time (sec) Group 1 -mM thiocholine Group 2 -mM thiocholine Group 3 -mM thiocholine Group 4 -mM thiocholine 30 60 90 120 150 180 210 240 270 300 Group 5 - A412 nm Group 6 - A412 nm Group 5 -mM thiocholine Group 6 -mM thiocholine...
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This note was uploaded on 10/18/2010 for the course BIO 204 taught by Professor O'neal during the Fall '07 term at SUNY Stony Brook.

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lab4datacollection_1 - 3.5 ml Use the control tube to blank...

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