Bio Lab 2 - 7. (see attached) 8. (see above) 9. Absorbance...

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Melissa Harintho Lab Section 1, Plantae South 1. I can assume that the affinity of Stanfordin for the IEC column matrix is less than NDase. 2. Potassium is the cation in the KOAc buffer that is exchanging with the NDase on the column matrix. 3. NDase has a greater overall net positive charge. I know this, because in the experiment, the elution point for Stanfordin was obtained before the elution point of NDase. This indicated that NDase had a greater affinity for the IEC column matrix, because it was more positively charged. 4. I would not achieve separation of Stanfordin and NDase in my sample if I used the 0.5 M concentration of KOAc buffer first, because a concentration of 0.5 M is strong enough to elute both Stanfordin and NDase at the same time. 5. The matrix in the IEC column I used in lab today has a net negative charge. I know this, because I exchanged a positive ion at the matrix interface. 6. a. b. (see attached)
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Unformatted text preview: 7. (see attached) 8. (see above) 9. Absorbance = 2.2749 Concentration: 0.226 (obtained in experiment) = 2.2749x x = 0.099345 mg/mL 0.099345 mg/mL = mg (protein in pool of NDase)/ 2.72 mL (obtained in experiment) 0.2702184 mg protein in pool of NDase 10. a. 33.773% b. (0.2702184 mg/0.8 mg) 100%= 33.733% c. 34.936% (average percent yield for lab section). My yield (33.773%) was slightly lower. This may have been due to slight experimental errors (i.e. dust on the spectrophotometer cuvets, carelessness in iteratively determining the volume of pooled NDase). d. Knowing the percentage yield in protein purification allows a molecular biologist to gauge the validity of her work. Percentage yield provides raw feedback about a biologists experimental procedure. Absorbance vs. Concentration y = 2.2749x R 2 = 0.9992 0.5 1 1.5 2 2.5 0.5 1 1.5 NDase concentration (mg/mL) Absorbance at 550 nm...
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