Anti-i-motif DNA mabs [iMbody](MABL-3601)

MABL-3601

BLI, confocal microscopy, ELISA, IF, Pull-down-seq

Engineer antibody

Species independent

iMbody

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

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Folded i‑motif DNA (conformational epitope)

The camelid‑derived nanobody iMbody was validated for its specificity against diverse i‑motif conformations by ELISA and confocal immunofluorescence assays. Although its exact dissociation constant (KD) has not been publicly reported, iMbody recognizes folded i‑motifs independent of primary DNA sequence, targeting intramolecular and intermolecular i‑motifs derived from i‑Myc, i‑RET, i‑hTelo, and i‑Nano sequences found within telomeric regions and proto‑oncogene promoters (Zeraati et al., 2023; DOI:10.1016/j.xpro.2023.102729). Under optimised experimental conditions, iMbody exhibits negligible cross‑reactivity toward off‑target nucleic‑acid structures, including G‑quadruplex (G4) DNAs (c‑MYC‑46, BCL2, KIT2, TBA, telomeric G4‑2JSM), double‑stranded DNA (dsDNA), DNA hairpins, and unstructured single‑stranded cytosine‑rich oligonucleotides. It has been successfully applied for high‑resolution immunofluorescence imaging to visualize endogenous i‑motif foci within HeLa cell nuclei, enabling sub‑nuclear localization analysis of i‑motif structures (Zeraati et al., 2023; DOI:10.1016/j.xpro.2023.102729). Furthermore, iMbody is compatible with pull‑down‑sequencing workflows to enrich and identify genomic i‑motif‑forming sequences under near‑physiological pH and temperature. Unlike the first‑generation iMab scFv antibody, there is no published evidence demonstrating that iMbody binding triggers unfolding of pre‑folded i‑motif structures. Nevertheless, its specificity remains sensitive to buffer composition, antibody concentration, and cell‑fixation‑permeabilization protocols. Partially‑folded or non‑canonical i‑motif variants retaining the conformational epitope may still be bound; therefore, stringent no‑nanobody blank controls and orthogonal cross‑validation alongside iMab, circular dichroism (CD) or NMR are strongly recommended to exclude false‑positive signals in experiments (Zeraati et al., 2023; DOI:10.1016/j.xpro.2023.102729).

Zeraati et al. Protocol for the production and purification of an i‑Motif‑specific nanobody. STAR Protoc. 2023 Dec 15;4(4):102729. doi: 10.1016/j.xpro.2023.102729. Epub 2023 Nov 22.

This article describes the generation and characterisation of the iMbody nanobody, as well as detailed workflows for its bacterial production, His‑tag purification, and practical applications including ELISA and immunofluorescence detection of i‑motifs within HeLa cell nucleiPMC

100 μg Purified antibody

lot of specific

Protein A affinity purified

PBS

Store at 4⁰C for up to 1 months. For longer storage, aliquot and store at - 20⁰C.