United Kingdom (UK)
Recombinant monoclonal antibody to DNA topoisomerase 1. Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from the chicken B-cell clone 5C2.
UniProt Accession Number of Target Protein: P11387
Alternative Name(s) of Target: TOP1; Topo; DNA topoisomerase I
Immunogen: The original antibody was generated by immunizing a barnevelder chicken with an antibody antigen cocktail comprising recombinant purified human antigens annexin XI, CENP-B, Topo, Jo-1, Ro60, Rpp38 and U1A in complete Freund's adjuvant. B cells that were harvested from the bone marrow and peripheral blood lymphocytes of immunized chickens was used to generate recombinant scFv phage display libraries. The specific clone was identified by panning the library against specific antigen.
Specificity: This antibody binds human topoisomerase 1. This enzyme controls and alters the topologic states of DNA during transcription. This enzyme catalyzes the transient breaking and rejoining of a single strand of DNA which allows the strands to pass through one another, thus altering the topology of DNA.
Application Notes: The binding characterization of this antibody was done using ELISA (PMID: 18062792).
Antibody first published in:
Hof et al. Multiple-antigen immunization of chickens facilitates the generation of recombinant antibodies to autoantigens. Clin Exp Immunol. 2008 Feb; 151(2): 367–377. PMID:18062792
Note on publication:
Describes the generation of chicked antibodies against various human autoantigens.
Immunofluorescence staining of HeLa cells with anti-DNA topoisomerase 1 antibody 5C2 (Ab03245). Immunofluorescence analysis of paraformaldehyde fixed HeLa cells, permeabilized with 0.15% Triton, on Shi-fix™ coverslips stained with the chimeric rabbit IgG version of 5C2 (Ab03245-23.0) (1:100 dilution) for 1 hour followed by Alexa Fluor® 488 secondary antibody (1:1000 dilution), showing nucleoli and cytoplasmic staining. The nuclear stain is DAPI (blue). Panels show, from left to right, top to bottom, Ab03245-23.0, DAPI, merged channels, and an isotype control. The isotype control was an unknown specificity antibody (Ab00178-23.0) followed by staining with Alexa Fluor® 488 secondary antibody.
Western blot using anti-DNA topoisomerase 1 antibody 5C2 (Ab03245). Jurkat cells lysate (35µg protein in RIPA buffer) was resolved via SDS-PAGE, and the subsequent blot was probed with the chimeric rabbit version of 5C2 (Ab03245-23.0) at 0.01µg/ml before detection using an anti-rabbit secondary antibody. A primary incubation of 1 hour was used, and proteins were detected by chemiluminescence.