Cat. No.
MABL-4032
Application
WB, ELISA
Isotype
Engineer antibody
Species Reactivity
E.coli
Clone No.
NT73
From
Recombinant Antibody
Specificity
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'.
Alternative Names
rpoC; tabB; epitope tag (SLAELLNAGLGGS); DNA-directed RNA polymerase subunit beta, RNAP subunit beta, RNA polymerase subunit beta, Transcriptase subunit beta
UniProt
P0A8T7
Immunogen
The original antibody was generated by immunizing BALB/c mice with recombinant ectodomain of glycoprotein E2 of GB virus, containing a double Strep-Tag of amino acid sequence 'AGWSHPQFEKGGGSGGGSGGGSWSHPQFEK'.
Application Notes
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.
Antibody First Published
Thompson et al. Isolation and characterization of a polyol-responsive monoclonal antibody useful for gentle purification of Escherichia coli RNA polymerase. Biochemistry (1992); 31(30):7003-8. PMID:1637835
Note on publication
Describes the generation and characterization of the antibody.
Size
100 μg Purified antibody.
Concentration
1 mg/ml.
Purification
Protein A affinity purified
Buffer
PBS with 0.02% Proclin 300.
Storage Recommendation
Store at 4⁰C for up to 3 months. For longer storage, aliquot and store at - 20⁰C.
