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SDS-PAGE analysis of the purification of Ub-Cas9. (A) Early-elution fractions. Lane 1, <t>E.</t> <t>coli</t> lysate; 2, flowthrough; 3, wash solution; E1–E3, eluted fractions with 80 mM imidazole; E4–E6, eluted fractions with 500 mM imidazole. (B) Late-elution fractions with 500 mM imidazole (E7–E15). For both panels, lanes B1–B5 contain 1–5 μg of BSA (asterisks) as a quantitative standard. The Ub-Cas9 protein is indicated by red arrowheads. M, protein standards.
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SDS-PAGE analysis of the purification of Ub-Cas9. (A) Early-elution fractions. Lane 1, <t>E.</t> <t>coli</t> lysate; 2, flowthrough; 3, wash solution; E1–E3, eluted fractions with 80 mM imidazole; E4–E6, eluted fractions with 500 mM imidazole. (B) Late-elution fractions with 500 mM imidazole (E7–E15). For both panels, lanes B1–B5 contain 1–5 μg of BSA (asterisks) as a quantitative standard. The Ub-Cas9 protein is indicated by red arrowheads. M, protein standards.
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SDS-PAGE analysis of the purification of Ub-Cas9. (A) Early-elution fractions. Lane 1, <t>E.</t> <t>coli</t> lysate; 2, flowthrough; 3, wash solution; E1–E3, eluted fractions with 80 mM imidazole; E4–E6, eluted fractions with 500 mM imidazole. (B) Late-elution fractions with 500 mM imidazole (E7–E15). For both panels, lanes B1–B5 contain 1–5 μg of BSA (asterisks) as a quantitative standard. The Ub-Cas9 protein is indicated by red arrowheads. M, protein standards.
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SDS-PAGE analysis of the purification of Ub-Cas9. (A) Early-elution fractions. Lane 1, E. coli lysate; 2, flowthrough; 3, wash solution; E1–E3, eluted fractions with 80 mM imidazole; E4–E6, eluted fractions with 500 mM imidazole. (B) Late-elution fractions with 500 mM imidazole (E7–E15). For both panels, lanes B1–B5 contain 1–5 μg of BSA (asterisks) as a quantitative standard. The Ub-Cas9 protein is indicated by red arrowheads. M, protein standards.

Journal: Bio-protocol

Article Title: A One-Step Method for Efficient Purification of Functional Cas9 Protein

doi: 10.21769/BioProtoc.5594

Figure Lengend Snippet: SDS-PAGE analysis of the purification of Ub-Cas9. (A) Early-elution fractions. Lane 1, E. coli lysate; 2, flowthrough; 3, wash solution; E1–E3, eluted fractions with 80 mM imidazole; E4–E6, eluted fractions with 500 mM imidazole. (B) Late-elution fractions with 500 mM imidazole (E7–E15). For both panels, lanes B1–B5 contain 1–5 μg of BSA (asterisks) as a quantitative standard. The Ub-Cas9 protein is indicated by red arrowheads. M, protein standards.

Article Snippet: E. coli DH5α cells (NEB, catalog number: C2988J) Reagents 1.

Techniques: SDS Page, Purification