Chemistry Laboratory Classwork

Useful Stats

Extinction Coefficient for HICA: 43430 M^-1cm^-1
Molar Mass for HICA: 26249.96 Da
A_(280nm)=ɛ_(HICA)bC_{(in M)} (account for dilution)
Mol/L to mg/mL
Use desalting volume of protein recovered to find mg of pure protein

Excel Data Files (dated and signed)

wt HICA Chromatograph Excel File: http://crystal-wiki.gonzaga.edu/tiki/tiki-download_file.php?fileId=148
R160H HICA Chromatograph Excel File: http://crystal-wiki.gonzaga.edu/tiki/tiki-download_file.php?fileId=149

[[Konis&Blount 1/28/13]]

*wt ECCA Chromatograph Excel File: http://crystal-wiki.gonzaga.edu/tiki/tiki-download_file.php?fileId=163

R160H HICA Chromatograph Excel File: http://crystal-wiki.gonzaga.edu/tiki/tiki-download_file.php?fileId=162

[[Labrum, Lee, & Bolinger, 3/4/13]]

*optimized R160H HICA Chromatograph: http://crystal-wiki.gonzaga.edu/tiki/tiki-download_file.php?fileId=201
[[Labrum, Lee, & Bolinger, 5/4/13]]

Summary Results

Proof of concept work (R160H)

Protein Variant	Elution Volume (mL)	Elution %B (imidazole concentration)	Yield in mg	total cell mass (g)	group member names
ECCA	34.93 mL	59%	2.84 mg	14.6 g	A. Labrum, D. Lee and I. Bolinger
HICA R160H	52.70 mL	55.6%	6.7 mg	13.1 g	A. Labrum, D. Lee, and I. Bolinger
HICA	42.3 mL	0%	0 mg	9.697 g	M. Konis, A. Blount
HICA R160H	batch gradient	100%	?	?	M. Konis, A. Blount

General Method

Variant	elution volume	elution %B or mM imidazole	yield protein (mg)	total cell pellet (g)	group members
T200H/S204H HICA		127 mM imidazole			J. Birge
M194H/T196H HICA	10.3 mL	105 mM imidazole	18.3 mg	5.60 g	T. Singh, J. Nguyen
M194H/T196H HICA	90.4 mL	100 mM imidazole	13.9 mg	6.61 g	J. Burr, N. Komlos
T200H/S204H HICA	9.03 mL	105 mM imidazole	0.0392 mg	5.62 g	J. Bauer, T. Heinrich
T200H/S204H HICA	12.1 mL	122 mM imidazole	10.2 mg	4.08 g	H. Horney, R. Zagal