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        <gnm:Cell Row="1" Col="1" ValueType="60">Ian Kleckner  -- Foster Lab -- ITC Sample Prep Calculator
v. 07/22/09
Fill in green boxes with your project-specific information.
Read out calculated values in yellow boxes.</gnm:Cell>
        <gnm:Cell Row="1" Col="2" ValueType="10"></gnm:Cell>
        <gnm:Cell Row="1" Col="3" ValueType="10"></gnm:Cell>
        <gnm:Cell Row="3" Col="2" ValueType="60">INPUT</gnm:Cell>
        <gnm:Cell Row="3" Col="3" ValueType="60">OUTPUT</gnm:Cell>
        <gnm:Cell Row="5" Col="1" ValueType="60">Name</gnm:Cell>
        <gnm:Cell Row="5" Col="2" ValueType="60">Ian Kleckner</gnm:Cell>
        <gnm:Cell Row="6" Col="1" ValueType="60">Date</gnm:Cell>
        <gnm:Cell Row="6" Col="2" ValueType="40" ValueFormat="m/d/yyyy">40013</gnm:Cell>
        <gnm:Cell Row="7" Col="1" ValueType="60">Notes</gnm:Cell>
        <gnm:Cell Row="7" Col="2" ValueType="60">Notes after completing first ITC experiment on 07/20/09
Got Kd2=0.257 uM for 2nd binding site and [TRAP] from E280 protparam</gnm:Cell>
        <gnm:Cell Row="8" Col="1" ValueType="60">Macromolecule</gnm:Cell>
        <gnm:Cell Row="9" Col="1" ValueType="60">Sample Name</gnm:Cell>
        <gnm:Cell Row="9" Col="2" ValueType="60">B. ste A26I TRAP</gnm:Cell>
        <gnm:Cell Row="10" Col="1" ValueType="60">Sample MW</gnm:Cell>
        <gnm:Cell Row="10" Col="2" ValueType="40">8242</gnm:Cell>
        <gnm:Cell Row="10" Col="3" ValueType="60">g/mol</gnm:Cell>
        <gnm:Cell Row="11" Col="1" ValueType="60">Sample notes</gnm:Cell>
        <gnm:Cell Row="11" Col="2" ValueType="60">HPLC purified, dialysis refolded on 07/10/2009</gnm:Cell>
        <gnm:Cell Row="12" Col="1" ValueType="60">Ligand</gnm:Cell>
        <gnm:Cell Row="13" Col="1" ValueType="60">Sample Name</gnm:Cell>
        <gnm:Cell Row="13" Col="2" ValueType="60">unlabeled tryptophan</gnm:Cell>
        <gnm:Cell Row="14" Col="1" ValueType="60">Sample MW</gnm:Cell>
        <gnm:Cell Row="14" Col="2" ValueType="40">204</gnm:Cell>
        <gnm:Cell Row="14" Col="3" ValueType="60">g/mol</gnm:Cell>
        <gnm:Cell Row="15" Col="1" ValueType="60">Sample notes</gnm:Cell>
        <gnm:Cell Row="15" Col="2" ValueType="60">Prepared from dialysate of pre-ITC TRAP</gnm:Cell>
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        <gnm:Cell Row="17" Col="4" ValueType="60">Recommendation (ik, 07/19/09)</gnm:Cell>
        <gnm:Cell Row="17" Col="5" ValueType="60">Notes</gnm:Cell>
        <gnm:Cell Row="18" Col="1" ValueType="60">Binding affinity Kd</gnm:Cell>
        <gnm:Cell Row="18" Col="2" ValueType="40" ValueFormat="##0.0E+00">2.57e-07</gnm:Cell>
        <gnm:Cell Row="18" Col="3" ValueType="60">M</gnm:Cell>
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        <gnm:Cell Row="18" Col="5" ValueType="60">CAM Kd=0.1uM</gnm:Cell>
        <gnm:Cell Row="19" Col="1" ValueType="60">Num binding sites (n)</gnm:Cell>
        <gnm:Cell Row="19" Col="2" ValueType="40">1</gnm:Cell>
        <gnm:Cell Row="19" Col="3" ValueType="60">-</gnm:Cell>
        <gnm:Cell Row="19" Col="4" ValueType="60">Usually 1</gnm:Cell>
        <gnm:Cell Row="21" Col="1" ValueType="60">Macromolecule concentration [M]</gnm:Cell>
        <gnm:Cell Row="21" Col="2" ValueType="40" ValueFormat="##0.0E+00">9.6606e-05</gnm:Cell>
        <gnm:Cell Row="21" Col="3" ValueType="60">M</gnm:Cell>
        <gnm:Cell Row="21" Col="4" ValueType="60">Varies</gnm:Cell>
        <gnm:Cell Row="22" Col="1" ValueType="60">c-value</gnm:Cell>
        <gnm:Cell Row="22" Col="2" ExprID="1">=C22*C20/C19</gnm:Cell>
        <gnm:Cell Row="22" Col="3" ValueType="60">-</gnm:Cell>
        <gnm:Cell Row="22" Col="4" ValueType="60">10 to 100</gnm:Cell>
        <gnm:Cell Row="22" Col="5" ValueType="60">CAM c=561 too high!</gnm:Cell>
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        <gnm:Cell Row="24" Col="2" ValueType="40">50</gnm:Cell>
        <gnm:Cell Row="24" Col="3" ValueType="60">-</gnm:Cell>
        <gnm:Cell Row="24" Col="4" ValueType="60">10 to 100</gnm:Cell>
        <gnm:Cell Row="25" Col="1" ValueType="60">Macromolecule concentration [M]</gnm:Cell>
        <gnm:Cell Row="25" Col="2" ExprID="2">=C25*C19/C20</gnm:Cell>
        <gnm:Cell Row="25" Col="3" ValueType="60">M</gnm:Cell>
        <gnm:Cell Row="25" Col="4" ValueType="60">Varies</gnm:Cell>
        <gnm:Cell Row="29" Col="1" ValueType="60">Ligand concentration [L]</gnm:Cell>
        <gnm:Cell Row="29" Col="2" ValueType="40" ValueFormat="##0.0E+00">0.000135</gnm:Cell>
        <gnm:Cell Row="29" Col="3" ValueType="60">M</gnm:Cell>
        <gnm:Cell Row="29" Col="5" ValueType="60">CAM used 662 uM</gnm:Cell>
        <gnm:Cell Row="30" Col="1" ValueType="60">Macromolecule concentration [M]</gnm:Cell>
        <gnm:Cell Row="30" Col="2" ValueType="40" ValueFormat="##0.0E+00">1.285e-05</gnm:Cell>
        <gnm:Cell Row="30" Col="3" ValueType="60">M</gnm:Cell>
        <gnm:Cell Row="30" Col="4" ValueType="60">n=1 concentration 71.295 uM</gnm:Cell>
        <gnm:Cell Row="30" Col="5" ValueType="60">CAM used 50 uM</gnm:Cell>
        <gnm:Cell Row="31" Col="1" ValueType="60">Num binding sites (n)</gnm:Cell>
        <gnm:Cell Row="31" Col="2" ValueType="40">1</gnm:Cell>
        <gnm:Cell Row="31" Col="3" ValueType="60">-</gnm:Cell>
        <gnm:Cell Row="31" Col="4" ValueType="60">Usually 1</gnm:Cell>
        <gnm:Cell Row="32" Col="1" ValueType="60">Molar binding enthalpy ΔH</gnm:Cell>
        <gnm:Cell Row="32" Col="2" ValueType="40">-12.42</gnm:Cell>
        <gnm:Cell Row="32" Col="3" ValueType="60">kCal/mol</gnm:Cell>
        <gnm:Cell Row="32" Col="5" ValueType="60">CAM dH=-13.88 kcal/mol
IK A26I dH=-12.42 kcal/mol</gnm:Cell>
        <gnm:Cell Row="34" Col="1" ValueType="60">Active cell volume</gnm:Cell>
        <gnm:Cell Row="34" Col="2" ValueType="40" ValueFormat="0.00">1.4166</gnm:Cell>
        <gnm:Cell Row="34" Col="3" ValueType="60">mL</gnm:Cell>
        <gnm:Cell Row="34" Col="4" ValueType="60">1.4166 mL</gnm:Cell>
        <gnm:Cell Row="35" Col="1" ValueType="60">Total volume titrated</gnm:Cell>
        <gnm:Cell Row="35" Col="2" ValueType="40" ValueFormat="0.00">285</gnm:Cell>
        <gnm:Cell Row="35" Col="3" ValueType="60">uL</gnm:Cell>
        <gnm:Cell Row="35" Col="4" ValueType="60">285 uL</gnm:Cell>
        <gnm:Cell Row="36" Col="1" ValueType="60">Injection volume</gnm:Cell>
        <gnm:Cell Row="36" Col="2" ValueType="40">5</gnm:Cell>
        <gnm:Cell Row="36" Col="3" ValueType="60">uL</gnm:Cell>
        <gnm:Cell Row="36" Col="4" ValueType="60">3 to 15 uL</gnm:Cell>
        <gnm:Cell Row="36" Col="5" ValueType="60">CAM 5 uL</gnm:Cell>
        <gnm:Cell Row="37" Col="1" ValueType="60">Injection spacing</gnm:Cell>
        <gnm:Cell Row="37" Col="2" ValueType="40">400</gnm:Cell>
        <gnm:Cell Row="37" Col="3" ValueType="60">sec</gnm:Cell>
        <gnm:Cell Row="37" Col="4" ValueType="60">At least 50 sec per uL injected</gnm:Cell>
        <gnm:Cell Row="39" Col="1" ValueType="60">Final molar ratio
[L] / [BindSites]</gnm:Cell>
        <gnm:Cell Row="39" Col="2" ExprID="3">=C36*C30/1000000/(C31*C32*C35/1000)</gnm:Cell>
        <gnm:Cell Row="39" Col="3" ValueType="60">-</gnm:Cell>
        <gnm:Cell Row="39" Col="4" ValueType="60">At least 1.5</gnm:Cell>
        <gnm:Cell Row="40" Col="1" ValueType="60">Num injections</gnm:Cell>
        <gnm:Cell Row="40" Col="2" ExprID="4">=C36/C37</gnm:Cell>
        <gnm:Cell Row="40" Col="3" ValueType="60">-</gnm:Cell>
        <gnm:Cell Row="40" Col="4" ValueType="60">At least 10</gnm:Cell>
        <gnm:Cell Row="41" Col="1" ValueType="60">Total experiment time</gnm:Cell>
        <gnm:Cell Row="41" Col="2" ExprID="5">=C38*C41/3600</gnm:Cell>
        <gnm:Cell Row="41" Col="3" ValueType="60">hrs</gnm:Cell>
        <gnm:Cell Row="42" Col="1" ValueType="60">Total heat evolved from first injection</gnm:Cell>
        <gnm:Cell Row="42" Col="2" ExprID="6">=C33*1000*C30*C37</gnm:Cell>
        <gnm:Cell Row="42" Col="3" ValueType="60">uCal</gnm:Cell>
        <gnm:Cell Row="42" Col="4" ValueType="60">Magnitude at least 3-5 uCal</gnm:Cell>
        <gnm:Cell Row="46" Col="1" ValueType="60">Num temperatures</gnm:Cell>
        <gnm:Cell Row="46" Col="2" ValueType="40">1</gnm:Cell>
        <gnm:Cell Row="46" Col="3" ValueType="60">temps</gnm:Cell>
        <gnm:Cell Row="46" Col="4" ValueType="60">At least 1</gnm:Cell>
        <gnm:Cell Row="47" Col="1" ValueType="60">Num repeats per temperature</gnm:Cell>
        <gnm:Cell Row="47" Col="2" ValueType="40">1</gnm:Cell>
        <gnm:Cell Row="47" Col="3" ValueType="60">repeats</gnm:Cell>
        <gnm:Cell Row="47" Col="4" ValueType="60">At least 1</gnm:Cell>
        <gnm:Cell Row="49" Col="1" ValueType="60">Volume of macromolecule per experiment</gnm:Cell>
        <gnm:Cell Row="49" Col="2" ValueType="40" ValueFormat="0.00">2.3</gnm:Cell>
        <gnm:Cell Row="49" Col="3" ValueType="60">mL</gnm:Cell>
        <gnm:Cell Row="49" Col="4" ValueType="60">2 mL</gnm:Cell>
        <gnm:Cell Row="50" Col="1" ValueType="60">Total volume of macromolecule</gnm:Cell>
        <gnm:Cell Row="50" Col="2" ExprID="7">=C50*C48*C47</gnm:Cell>
        <gnm:Cell Row="50" Col="3" ValueType="60">mL</gnm:Cell>
        <gnm:Cell Row="51" Col="1" ValueType="60">Total macromolecule required</gnm:Cell>
        <gnm:Cell Row="51" Col="2" ExprID="8">=C51/1000*C31*C11*1000</gnm:Cell>
        <gnm:Cell Row="51" Col="3" ValueType="60">mg</gnm:Cell>
        <gnm:Cell Row="53" Col="1" ValueType="60">Volume of ligand per experiment</gnm:Cell>
        <gnm:Cell Row="53" Col="2" ValueType="40" ValueFormat="0.00">5</gnm:Cell>
        <gnm:Cell Row="53" Col="3" ValueType="60">mL</gnm:Cell>
        <gnm:Cell Row="53" Col="4" ValueType="60">0.5 mL</gnm:Cell>
        <gnm:Cell Row="54" Col="1" ValueType="60">Total volume of ligand</gnm:Cell>
        <gnm:Cell Row="54" Col="2" ExprID="9">=C54*C48*C47</gnm:Cell>
        <gnm:Cell Row="54" Col="3" ValueType="60">mL</gnm:Cell>
        <gnm:Cell Row="55" Col="1" ValueType="60">Total macromolecule required</gnm:Cell>
        <gnm:Cell Row="55" Col="2" ExprID="10">=C55/1000*C30*C15*1000</gnm:Cell>
        <gnm:Cell Row="55" Col="3" ValueType="60">mg</gnm:Cell>
      </gnm:Cells>
      <gnm:MergedRegions>
        <gnm:Merge>C16:E16</gnm:Merge>
        <gnm:Merge>C14:E14</gnm:Merge>
        <gnm:Merge>C12:E12</gnm:Merge>
        <gnm:Merge>C10:E10</gnm:Merge>
        <gnm:Merge>C8:E8</gnm:Merge>
        <gnm:Merge>C7:E7</gnm:Merge>
        <gnm:Merge>C6:E6</gnm:Merge>
      </gnm:MergedRegions>
      <gnm:SheetLayout TopLeft="B31"/>
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  <gnm:Calculation ManualRecalc="0" EnableIteration="1" MaxIterations="100" IterationTolerance="0.001"/>
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