Isothermal Titration Calorimetry

Sample Submission Form

Complete a separate request form for each sample

Contact Information

Data.

Do you want a hard copy of your data? Note: All data will also be emailed to the email address provided, above.

No hard copy is necessary.

Yes, please send a hard copy of my data.

Address for hard copy.

Experimental Details

Expectations. This information helps us to offer advice on experimental design.

Acquisition Parameters. Please enter the data acquisition parameters based on your knowledge of the sample.

Temperature (°C).

Other instructions.

Sample Guidelines & Information.

We accept NO radioactive samples.

The minimal guidelines below must be followed to ensure an accurate estimate of stoichiometry (n), heat of binding (DelH), and binding constant (Kb) (or dissociation constant Kd = 1/Kb).

• Sample

  • Volume at least 1.5 ml.

  • The lowest concentration which can be studied is 3 mM and this is adequate only for tight binding where Kd is smaller than 1 mM. For weaker interactions, the macromolecule concentration should be 5 times Kd, or higher if possible. Preferably, the macromolecule solution should be dialyzed exhaustively against buffer for final equilibration.

• Ligand (the sample to be placed in the injection syringe).

  • Volume of at least 0.7 ml.

  • Concentration at least 10 times higher than the macromolecule concentration.

  • If the macromolecule has multiple binding sites for ligand, then the ligand concentration must be increased accordingly.

• Buffer. The buffer solution in which the ligand is dissolved should be exactly the same buffer against which the macromolecule has been equilibrated.

  • Dialyze the sample solution exhaustively against buffer.

  • Provide at least 20 ml of buffer.

• The pH of each should be checked carefully after both solutions have been prepared. If they are different by more that 0.05 pH units, then one of the solutions must be back-titrated so they are within the limit of 0.05 pH units.

• If any solution contains visible particles, they should be filtered out.

• If possible, the concentrations of both solutions should be accurately determined after final preparation. Accurate determination of binding parameters is only possible if concentrations of binding components are known precisely on the final solution.

• DTT should be avoided and replaced by b-mercaptoethanol or TCEP.

SAMPLE

SAMPLE name, composition, and concentration.

Your SAMPLE ID number.

LIGAND.

LIGAND name, composition, and concentration.

Your LIGAND ID number.

Molelcular Weight		Volume (ml)
pH

BUFFER.

BUFFER name, composition, and concentration.

Your BUFFER ID number.

Volume (ml)		pH

ESTIMATES.

Estimation of Ka (include units).

Estimation of stoichiometry (Ligand:Sample, 1:1, 2:1, etc).

SAMPLE DELIVERY.

When will the sample will be ready for us?

Explain any special handling for your sample.

You can print the next verification page after you submit your request. We will contact you to schedule your request. If you do not hear from us within 48 hours, please contact us.