Anti-i-motif DNA mAbs [iMab](MABL-1916)

MABL-1916

BLI, confocal microscopy, ELISA, IF

Engineer antibody

Species independent

iMab

Recombinant Antibody

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.

intercalated motif; i-motif; intercalated DNA motif; I motif; DNA i-motif; human telomere i-motif ; hTelo i-motif; imotif; imotiv; imotive; i-motiv; i-motive

The original antibody was selected from the Garvan-2 human scFv library via phage display.

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 imm 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 antibody's selectivity, emphasizing the importance of optimizing DNA concentrations, blocking conditions, and washing protocols. These factors are crucial for confirming the antibody's selectivity for i-motifs and ensuring accurate detection of both intramolecular and intermolecular i-motifs (Ruggiero et al., 2024; DOIunofluorescence 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: 10.1101/2024.06.22.600195v1).

Zeraati et al. I-motif DNA structures are formed in the nuclei of human cells. Nat Chem. 2018 Jun;10(6)：631-637. doi： 10.1038/s41557-018-0046-3. Epub 2018 Apr 23. PMID：29686376

This article describes the generation and characterisation of the iMab antibody fragment and its use in the detection of i-motifs in the nuclei of human cells.

100 μg Purified antibody.

1 mg/ml.

Protein A affinity purified

PBS with 0.02% Proclin 300.

Store at 4⁰C for up to 3 months. For longer storage, aliquot and store at - 20⁰C.