Figure 1.

Purification of LigIV Constructs

(A) Profile of the UV absorbance at 280 nm during heparin affinity chromatography. The absorbance during heparin is shown in blue. Arrows indicate the ranges of fractions used for SDS-PAGE.

(B) SDS-PAGE gel of fractions eluted from a heparin column. The molecular weight markers are in column “M” and the molecular weights (kDa) of the gel are shown on the left of the gel. Protein bands are indicated on the right of the gel: LigIV^1–609 (residues 1–609 of LigIV), BRCT (residue 645–911 of LigIV) and XRCC4 (residue 1–213 of XRCC4, all cysteines of which are mutated to alanines).

(C) Profile of the UV absorbance at 280 nm during size exclusion chromatography. The absorbance during size-exclusion chromatography is shown in red.

(D) SDS-PAGE gel of fractions eluted from a Superdex 200 column. The molecular weight markers are identical to those shown in (B). Protein bands are indicated on the right of the gel.

(E) SDS-PAGE gels of purified LigIV^1–609. A total of 2 μg of both LigIV^1–609 (left) and LigIV^1–620 (right) are shown with their molecular weights (kDa).

See also Figure S1.