CHEM 271 Lecture Notes - Lecture 4: Absorbance, Vitamin C, Elution
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Ldh activity not calculated after the 5th fractional number. Though in theory it should be negligible or close to zero. Absorbance of cytochrome c (at = 550 nm ) Absorbance of fmn (at = 450 nm ) % ldh activity of fraction 5: 0. 011*100%/1. 117 = 9. 8% % change of fmn: 100 [(100*1. 8000)/1. 8232] = 1. 27% % change of cytochrome c: 100 [(100*1. 1067)/0. 7920] = 39. 73: in what order did you expect fmn, ldh and cytochrome c to elute from the column. One would expect the order of elution to be ldh, cytochrome c, and finally fmn. This is due to gel filtration chromatography eluting is order of size, ldh being the largest at 140000g/mol followed by. It was observed that sample 5 of elute was clear, which represents the ldh activity, followed by. If we were to not assay the ldh activity, there would be no way to detect when the ldh would elute.
