Skip to main content
. Author manuscript; available in PMC: 2009 Apr 1.
Published in final edited form as: Protein Expr Purif. 2007 Dec 4;58(2):184–193. doi: 10.1016/j.pep.2007.11.010

Figure 1. Mouse recombinant his-ACBP cloning and purification.

Figure 1

A. Agarose gels of the his-ACBP PCR product, his-ACBP PCR fragment cloned into pGEM-T vector, and his-ACBP subcloned into pET21b vector: HMW, high molecular weight marker (1kb); LMW, low molecular weight marker (100bp); H-ACBP-PCR, PCR product encoding his-ACBP cDNA; pGEM-H-ACBP, his-ACBP PCR fragment T-A cloned into the pGEM-T plasmid: 1, 1μg of uncut plasmid; 2, 1μg of Xho I/Nde I cut plasmid; pET21b-H-ACBP refers to his-ACBP cDNA cloned into pET21b plasmid: 3, 1μg of Xho I -cut plasmid; 4, 1μg of Xho I/Nde I digested plasmid. B. DNA sequence of mouse ACBP cDNA inserted into pET21b vector and its amino acid translation. C. SDS-PAGE gel and Western blotting of his-ACBP expressed in BL21 E. coli cells in the absence (lanes 1 and 3, respectively) and presence of IPTG (lanes 2 and 4, respectively). For SDS-PAGE and Coomassie staining, 20 μg of total protein was loaded per lane, while 10 μg of total protein per lane was used for Western blotting. D. Representative Coomassie stained SDS-PAGE gel showing his-ACBP in E. coli cell lysate (lane 1, 25 μg protein) and elution fraction after Ni-column (lane 2, 2 μg protein). SDS-PAGE gel of purified his-ACBP silver stained to determine purity: 3, protein marker; 4, ultra low range protein marker; 5, 1μg of his-ACBP; 6, 2μg of his-ACBP; 7, 5μg of his-ACBP.