Figure 1 | British Journal of Cancer

Figure 1

From: Isolation and characterisation of a human anti-idiotypic scFv used as a surrogate tumour antigen to elicit an anti-HER-2/neu humoral response in mice

Figure 1

Analysis of antibody-containing periplasmic fractions containing antibodies (A) After induction of single ampicillin-resistant infected E. coli HB2151 colonies with 1 mM IPTG. Lane 1: clone 39; lane 2: clone 40; lane 3: clone 92; lane 4: clone 69. After size fractionation on 12.5% SDS–PAGE, protein extracts were blotted onto nitrocellulose. The immunoblot was developed with HRP-conjugated M2 anti-FLAG mAb (1 : 2000) followed by addition of the 4-chloro-1-naphtol substrate. (B) Before and after purification. The purity was controlled on 12.5% SDS–PAGE gel followed by silver staining. Lane a: nonpurified periplasmic fraction. Lane b: Periplasmic fraction purified on a Hitrap Ni-activated chelating column. Lane c: Periplasmic fraction purified on a Hitrap Ni-activated chelating column followed by gel filtration on Superdex 75. Lane d: Standard molecular mass markers.

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