Protein Purification Practice Problem

Protein Purification Practice Problem - 1. Pepsin, His...

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Protein Purification Instructions: In your group, design a possible purification protocol for one of the combinations below. For each step, identify the tool you are using, any special buffer conditions , and the order you expect things to elute. Be prepared to present your protocol to the class. Your tools: Size exclusion chromatography (15-100 kDa resin) Anion exchange Cation exchange Ligand affinity chromatography Metal binding chromatography UV Detectors SDS-PAGE gel electrophoresis Mixture:
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Unformatted text preview: 1. Pepsin, His 6-Ube2t, progesterone receptor, BRCA1 2. Ubiquitin, insulin, His 6-Ube2t, hemoglobin 3. BRCA1, hemoglobin, pepsin, insulin 4. Hemoglobin, myoglobin, insulin, progesterone receptor Protein Molecular Weight (kDa) pI Pepsin (pre-cleavage/activation) 34.6 3.36 Insulin 12.0 5.22 Myoglobin 17.0 7.29 Hemoglobin Dimer of heterodimers 7.77 (HbA) (15.1) (8.73) (HbB) (15.9) (6.81) His 6-Ube2t 25.8 6.44 Progesterone Receptor 29.5 8.42 BRCA1 207.7 5.29 Ubiquitin 8.5 6.56...
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