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Figure 2 | BMC Biochemistry

Figure 2

From: Streptavidin-Binding Peptide (SBP)-tagged SMC2 allows single-step affinity fluorescence, blotting or purification of the condensin complex

Figure 2

Analysis of single-step purification of SBP-SMC2. (A) Equivalent amounts of protein eluent from untagged DT40 wild type and SMC2-SBP preparations were analysed by silver staining. SMC2-SBP and its condensin dimerisation partner SMC4 appear as the predominant bands in tagged eluent. (B) Standardised protein amount of crude input (I) and eluent (E) of both untagged and SMC2-SBP were immunoblotted using rabbit anti-SMC2 antibody followed by anti-rabbit-AP to detect SMC2 proteins. SMC2-SBP is present in both E and I, whilst wild-type (WT) SMC2 is not present after purification. (C) Single step affinity blotting of SMC2-SBP. The same samples in (B) were affinity blotted using streptavidin-AP. SMC2-SBP is detected in both input, and eluent of the SMC2-SBP tagged extracts, whilst, not in untagged extracts. Streptavidin contaminants 1 and 2 are present in both untagged and tagged. Note indicates SMC2 degradation product, and * indicates SMC2-SBP detected by streptavidin-AP.

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