Calculate the molar concentration of substrate

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SUMMARY

The discussion focuses on calculating the molar concentration of the substrate Paranitrophenylphosphate for a Michaelis-Menten curve. The substrate concentration is given as 1 mg/ml, and the formula weight is 275 g/mole. The user successfully converted the volume of substrate (10 µL) to mass and then to moles, ultimately calculating a molarity of 3.580e-4 M for the substrate in a 100 µL solution. The methodology involves converting volumes to mass, mass to moles, and then determining concentration in molarity.

PREREQUISITES
  • Understanding of molarity and its calculation
  • Knowledge of the Michaelis-Menten kinetics
  • Ability to perform unit conversions (mL to L, mg to g)
  • Familiarity with the concept of moles and molar mass
NEXT STEPS
  • Learn about calculating molar concentration using different substrates
  • Explore the Michaelis-Menten equation and its applications in enzyme kinetics
  • Study the effects of substrate concentration on reaction rates
  • Investigate common laboratory techniques for preparing dilutions
USEFUL FOR

Students in biochemistry, laboratory technicians, and researchers involved in enzyme kinetics and substrate concentration calculations will benefit from this discussion.

Thrax
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Homework Statement


Hello,
I'm creating a Michealis/Menten curve and need to find the X values for the graph.


Homework Equations



These are the instructions given to find the X values:

To obtain the X value, plot the molar concentration (molarity) of substrate used in each well.

You can determine this by multiplying the volume of substrate added to each well by the concentration of the substrate (1 mg/ml or 1 ug/ul) to give you a mass.


Paranitrophenylphosphate (the substrate) has a formula weight of 275 g/mole.

The substrate is diluted in 100uL in each well.

Using these values, calculate the molar concentration of substrate in each well across the plate. These are the X values. You are now ready to plot the kinetics curve.

The Attempt at a Solution



Using one data set of for 10uL of substrate i calculated moles
First converted the 10uL to .010mL

.010mL * 1 mg/ml = .010mg mass of substrate

Converted the .010mg to grams and solved for moles

1.000 x10-5g (1mol / 279g) = 3.58 x10e-8 moles

converted the 100ul to liters

100e-6uL = 1.000e-4 L


then found found moles\liters

3.58 x10e-8 moles / 1.000e-4 L = 3.580e-4 M

Not sure if this is correct or not. I also tried taking the 10ug / 100ul = .100 ug/uL and doing something with that, but got totally lost...
 
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I have just skimmed not checking numbers, but it looks to me like you are following the right path - volume to mass, mass to moles, moles to concentration.


--
 
Thanks for taking a look!
 

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