Spectrabind Protocol

1. For each experiment that you complete make a text file with four columns and be sure to include your zero point. Colums are set as indicated below:

Anisotropy Non-volume corrected total intensity [Oligo] [Protein]

2. Save the file as a text file with space delimited columns (.prn extension) and e-mail the file to yourself

3. Insert your Spectrabind zip disk with all the necessary files and download your data onto the zip disk

4. Open the file <Howmany.dat> and make sure numbers are correct:

a. Number of species = 3

b. Number of equations = 1

c. Number of elements = 2

d. Number of species = 2

e. Number of samples = number of titration points (including the zero point)

5. Save the file

6. Open the file <kguesses.dat>

7. Make sure the initial guess for the dissociation constant is <+1e+0>

8. Save the file

9. Open the file <Keformat.dat>

10. Enter the number of titration points

11. Enter <2> as the number of elements

12. Enter the file name of the text file with your data

13. Enter .001 as absolute error for anisotropy

14. Enter .001 as absolute error for intensity

15. Enter .01 as relative error for element 1

16. Enter .01 as relative error for element 2

17. Once the computer is done working close the window

18. Open file <Spctranew.dat>

19. Once the computer is done working close the window

20. Open the file <Kbestfit.txt>

21. Record the KBestFit and the range of values given in the brackets

22. Open the file <Spec001.txt> and record the two values for species 1 (not the error range)

23. Open the file <Spec002.txt> and record the two values for species 2 (not the error range)

24. Open the file <Localchi.txt> and look at the numbers to see if they are sufficiently low (<2)

25. Repeat Steps 4-25 for each experiment

26. When finished Close all the windows, leave the computer on, turn off the monitor