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Calculator
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Discussion
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How it is
done
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A. Isotope
decay
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Calculates radioactive decay during a
specified number of days. Select one of the common
isotopes, or enter the half-life of another isotope.
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The result becomes one of the
inputs to calculator B.
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The
decay constant (k) equals 0.693/HalfLife. The fraction
remaining equals e-kt.
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B. Conc. of
stock
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Enter
the original mCi/ml and Ci/mmole, which should be on the
label. If you are using a molecule labeled with 125I, the
specific activity equals 2200 Ci/mmole if each molecule is
labeled with one iodine.
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Also enter the percent of the
original isotope remaining (calculated in A
above). The calculations assume that the decay
product is not biologically active, so the
concentration of biologically active stock
decreases over time.
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The result is used as one of the
inputs to calculator C.
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Divide the original mCI/ml by Ci/mmole to
get the original concentration in mmole/ml which is the
same as molar. Multiply by the fraction remaining to get
the current concentration.
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C. Dilution of
stock
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Enter
the concentration in your stock solution, after accounting
for decay (calculated in calculator B). Also enter the
concentration and volume you want.
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The result is the volume of
stock you need to use.
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Divide the desired concentration by the
stock concentration to get the dilution factor. Multiply
that times the volume desired to get the volume of stock
needed.
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D. Specific activity
(cpm/fmol)
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Enter
the specific radioactivity as Ci/mmole which should be on
the label. If you are using a molecule labeled with 125I,
the specific activity equals 2200 Ci/mmole if each molecule
is labeled with one iodine.
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Also enter the counter
efficiency - the fraction of radioactive
disintigrations that are detected. The efficiency
depends on the isotope and instrumentation. With
low energy isotopes such as tritium, the
efficiency also depends on the experimental
details such as the choice of scintillation fluid,
the amount of water in the sample, and the
presence of any colored substances in the
sample.
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The calculation does not account
for radioactive decay. We assume that the decay
products are not biologically active. As the
molecules decay, the concentration goes down, but
the number of cpm per fmol of active molecule does
not change.
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The result is used as inputs to
calculators E, F and G
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A
Curie equals 2.2 x 1012 radioactive disintigrations per
minute (dpm). Multiply Ci/mmole by that factor to get dpm
per mmole. Multiply by the efficiency to get cpm per mmole,
and divide by 1012 to get cpm/fmol.
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E. Cpm to
fmol/mg
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Enter
the specific radioactivity as cpm/fmol, the number of cpm
counted, and the protein content of the sample in mg. The
result is the number of binding sites in fmol/mg
protein.
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Divide cpm by cpm/fmol to determine the
number of fmols. Divide by mg to get fmol/mg.
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F. Cpm to
sites/cell
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Enter
the specific radioactivity as cpm/fmol, the number of cpm
counted, and the cell count . The result is the number of
binding sites per cell.
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Divide cpm by cpm/fmol to get the number
of fmols. Multiply by 6.02 x 10 8 to get the number of
molecules. Divide by cell number.
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G. Cpm to nM
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Enter
the specific radioactivity as cpm/fmol, the number of cpm
counted, and the volume counted. The result is the
concentration in nM.
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Divide cpm by cpm/fmol to get the number
of fmols. Divide by volume.
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