Cat. No.
MABL-3254
Application
EMSA, gel retardation assay, IB, in vitro, SPR, ELISA, FC, IF, IHC
Isotype
Engineer antibody
Species Reactivity
Species independent
Clone No.
Z22
From
Recombinant Antibody
Specificity
Z22 binds both Z-DNA, but not B-DNA or ssDNA (single-stranded DNA). It recognizes Z-DNA at the phosphodiester backbone of various base sequence including (dG-dC)n.(dG-dC)n, (dTdG)n.(dC-dA)n, (dG-dme5C)n.(dG-dme5C)n and (dG-dbr5C)n.(dG-dbr5C)n. In other words, Z22 binds Z-DNA irrespective of sequence. Z22 Also binds DNA-8-MOP aducts and Z-RNA.
Alternative Names
Z-22; Z 22; Z DNA; ZDNA; Z RNA; ZRNA; Z NA; Z-NA; ZNA
UniProt
Immunogen
Z22 was prepared by immunizing C57BL/6 mice with brominated poly(dG-dC).poly(dG-dC) complexed with methylated bovine serum albumin (BSA), and was selected for by ELISA. Brominated poly(dG-dC).poly(dG-dC) forms a stable Z-DNA helix under physiological salt conditions.
Application Notes
Z22 binding to Z-DNA was evaluated by a competitive solid-phase ELISA (Möller et al., 1982; PMID: 7118931). The binding affinity of Z22 Fab version engineered from the original mouse IgG2b format to Z-DNA by SPR was measured to have an apparent KD of ~160 nM and could be competed out by soluble brominated d(G-C)15 but not unmodified d(G-C)15 (B-DNA). Z22 scFv binds to the target with a similar affinity to Z22 Fab. Competitive ELISA was used to determine binding specificity. Gel retardation assays were also used to show Z22 binding to DNA-8-MOP adducts. Biotinylated Z22 was used to determine the distribution of Z-DNA in permeabilized, microbead-encapsulated nuclei after adding radioactive streptavidin, and compared to encapsulated permeabilized nuclei stained with DAPI (Wittig et al., 1989; PMID: 2921282). Z22 was shown to cross-react with Z-RNA (Zarling et al., 1990; PMID: 2153833). Z-RNA binding using this product (Ab00783-3.0) was shown using IF (Zhang et al., 2020; PMID: 32200799), and it was also used in FC (Koehler et al., 2021; PMID: 34192517). Furthermore, this product (Ab00783, unspecified isotype) was also used in IHC and IB (Yau et al., 2021; PMID: 34728780), and Electrophoretic Mobility Shift Assay (EMSA; gel mobility assay) (Li et al., 2022; PMID: 35744832).
Antibody First Published
Möller et al. Monoclonal Antibodies Recognize Different Parts of Z-DNA. J Biol Chem. 1982 Oct 25;257(20):12081-5. PMID:7118931
Note on publication
Describes the production of monoclonal antibodies which recognize Z-DNA. Binding specificity and affinity were determined.
Size
100 μg Purified antibody.
Concentration
1 mg/ml.
Purification
Protein A affinity purified
Buffer
PBS with 0.02% Proclin 300.
Storage Recommendation
Store at 4⁰C for up to 3 months. For longer storage, aliquot and store at - 20⁰C.
