CloneID: DNA-1
Heavy Chain modification: Fc Silent™
Antigen Long Description: The original format of this antibody was isolated from a phage display derived from MRL/lpr mice (autoimmune mice). The library was panned against single-stranded calf thymus DNA.
Buffer Composition: PBS only.
Chimeric Use Statement: This chimeric rabbit antibody was made using the variable domain sequences of the original Mouse IgG1 format, for improved compatibility with existing reagents, assays and techniques.
Specificity Statement: This antibody is specific for dT5, with high specificity to sequences greater than 5 nucleotides. It primarily recognizes the thymine DNA base, but it also binds to oligo(dT)15 nucleotides or greater and oligo(dT) of length three to five nucleotides. The antibody also recognizes the abbreviated hairpin sequence LIG5-14.
Application Notes (Clone): The original format of this antibody (Fab) was generated and characterized. It was shown to bind DNA in an IP assay. and its binding affinity was determined to be (4.4±2.4) × 10-8 M (Calcutt et al., 1993; PMID: 7506692). Fluorescence titration of the antibody with dT5 was performed, and a Kd of ~27 µM was measured (Komissarov et al., 1996; PMID: 8647821). The crystal structure of the Fab fragment in complex with dT5 was determined (Tanner et al., 2001; PMID: 11733999). Isothermal titration calorimetry experiments were used to measure the apparent affinity of the antibody for dT5. The apparent association constant was KA = 6.0 × 104 M-1. Furthermore, the binding of dT5 to the antibody was exothermic, with an apparent ΔHcal under solution conditions of -16.8 kcal/mol (Schuermann et al., 2004; PMID: 15340914). The crystal structure of the Fab version of this antibody was characterized in complex with a DNA ligand corresponding to nucleotides 5–14 of LIG1-17 (LIG5-14) (Ou et al., 2007; PMID: 18028946).
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