CloneID: NT73
Buffer Composition: PBS only.
Uniprot Accession No.: P0A8T7
Specificity Statement: This antibody recognizes and binds an epitope located at the C terminus of beta subunit of the Escherichia coli RNA polymerase. It specifically binds a 13-amino-acid long sequence 'SLAELLNAGLGGS'.
Application Notes (Clone): This antibody was initially generated for purification of Escherichia coli RNA polymerase because of its ability to release the RNA polymerase in the presence of a low molecular weight polyhydroxylated compound (polyol) and a non-chaotropic salt. Such antibodies were termed as "polyol responsive" MAbs. Using NT73 conjugated to Sepharose, highly active RNA polymerase could be prepared rapidly by a single immunoaffinity chromatography step, replacing two lengthy chromatographic steps in the conventional purification procedure. The polyol-responsiveness of the antibody was determined by an ELISA-elution assay (PMID: 1637835). Later on the epitope of the abtibody was recognized and found to be a 13-amino-acid sequence 'SLAELLNAGLGGS' which was converted to an epitope tag that can be fused to a protein of interest for use as a purification tag. This epitope tag called 'Softag1' was fused to either the N or the C terminus of the green fluorescent protein. These tagged proteins were expressed in E. coli, and the tagged proteins were purified from the soluble fraction by a single-step immunoaffinity chromatography procedure. This approach extends the powerful technique of gentle-release immunoaffinity chromatography to many expressed proteins (PMID: 14656522). The scFv variants of this antibody loses considerable affinity for its antigen, however it maintains similar polyol-responsiveness as the parent monoclonal antibody (PMID: 16216525).
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