Anti-i-motif DNA [iMab]

Absolute Antibody

Anti-i-motif DNA [iMab]

7

This antibody is specific for the DNA i-motif, a four-stranded DNA structure that forms in cytosine-rich regions and is potentially involved in regulating DNA function.

The original human scFv version of this antibody was validated for its specificity towards various i-motif types through ELISA and bio-layer interferometry (BLI), with a dissociation constant (KD) of 59 nM. It selectively binds to i-motifs, including those in 1G22, 1A83, c-MYC, RET, and VEGF sequences, located in regions such as telomeres, centromeres, and proto-oncogene promoters. Importantly, this antibody does not interact with structures resembling i-motif DNA, such as DNA G4 quadruplex forms, double-stranded DNA (dsDNA), hairpin DNA, miRNA, or mutant human telomeres lacking i-motifs (Zeraati et al., 2018; PMID: 29686376). Notably, it does not bind to various DNA G4 quadruplex forms, including promoter G4s c-MYC44, BCL245, VEGF, telomeric 2JSM47, 2GKU, or aptamer G4 TBA. Additionally, the antibody has proven effective for immunofluorescence detection of i-motifs in multiple cell lines (MCF7, U2OS, and HeLa), confirming the presence of i-motif structures in human nuclei and their dependence on pH and cell-cycle stage (Zeraati et al., 2018; PMID: 29686376). This antibody selectively binds to both intramolecular and intermolecular i-motif structures. The composition of buffers used during binding and washing steps significantly influences the antibodys selectivity, emphasizing the importance of optimizing DNA concentrations, blocking conditions, and washing protocols. These factors are crucial for confirming the antibodys selectivity for i-motifs and ensuring accurate detection of both intramolecular and intermolecular i-motifs (Ruggiero et al., 2024; DOI: 10.1101/2024.06.22.600195v1).

ImmunoFluorescence, ELISA, Other Application

ELISA; BLI; IF; confocal microscopy

Research Use Only

Monoclonal

iMab

Mouse

IgG2a

This chimeric mouse IgG2a antibody was made using the variable domain sequences of the original scFv format, for improved compatibility with existing reagents, assays and techniques.

Other Species

This antibody fragment was selected via phage display from the Garvan-2 human single-chain variable fragment (scFv) library.

-20°C,2°C to 8°C

12352203