Anti-Zinc Fingers Linker (ZnFL) Antibody

C2H2 zinc finger (ZNF) proteins (ZFPs) represent the largest family of gene regulators in human. C2H2 ZFPs wrap around DNA via multiple zinc fingers to ensure a stable DNA interaction. Small linker peptides joining adjacent zinc finger modules are critical for the DNA-binding activity of C2H2 ZFPs. The vast majority of linker peptides contain a serine or threonine as their first amino acid residue, phosphorylation of which greatly reduces the DNA binding activity of C2H2 ZFPs. About half of ZFPs contain the highly conserved KRAB repression domain. KRAB family zinc finger proteins (KRAB-ZNF) adopt simple protein architecture where the conserved N-terminal KRAB domain is linked with the sequence-specific DNA-binding domain comprised of tandem arrays of the C2H2 type ZNFs. The KRAB domain directly binds KAP1 and is essential for KRAB-ZNF-mediated gene repression. Upon recruitment to chromatin by KRAB-ZNFs, KAP1 in turn functions as a scaffolding protein for histone- and DNA-modifying enzymes involved in target genes silencing.

~20-180 kDa observed. Variable depending on the sizes of the zinc finger proteins present in samples. Uncharacterized band(s) may appear in some lysates.

A mixture of zinc fingers linker (ZnFL) consensus peptides (Addison, J.B., et al. (2015). Cancer Res. 75(2):344-355).

Epitope: Zinc fingers linker (ZnFL).

Immunoprecipitation Analysis: A representative lot immunoprecipitated endogenous KRAB-ZNFs from human MDA-MB-231LN breast cancer cells and non-cancer HMLE mammary epithelial cells (Addison, J.B., et al. (2015). Cancer Res. 75(2):344-355).

Western Blotting Analysis: A representative lot detected ZnFL-specific target bands in the nuclear fractions from all vertebrate samples tested, although only three weak ZnFL-reactive bands were detected in chicken cell lysates. This antibody did not detect ZNFs in yeast and reacted with only one band in E. coli and a few proteins in Drosophila cell lysates (Addison, J.B., et al. (2015). Cancer Res. 75(2):344-355).

Anti-HpTGEKP motif, Cat. No. ABE319, is also available for detecting zinc finger linker threonine phosphorylation in Flow cytometry, Immunocytochemistry, Immunoprecipitation, and Western blotting applications.

Research Category
Epigenetics & Nuclear Function

Research Sub Category
Transcription Factors

This Anti-Zinc Fingers Linker Antibody is validated for use in Western Blotting, Immunoprecipitation for the detection of ZnFL.

This polyclonal antibody detected strong ZnFL-specific signals in the nuclear fractions from all vertebrate cells tested, although only three weak ZnFL-reactive bands were detected in chicken cell lysates. This antibody did not detect ZNFs in yeast and reacted with only one band in E. coli and a few proteins in Drosophila cell lysates (Addison, J.B., et al. (2015). Cancer Res. 75(2):344-355).

Affinity purified.

Purified rabbit polyclonal antibody in PBS with 40% glycerol and 0.05% sodium azide.

Stable for 1 year at -20°C from date of receipt.
Handling Recommendations: Upon receipt and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.
Note: Variability in freezer temperatures below -20°C may cause glycerol containing solutions to become frozen during storage.

Evaluated by Western Blotting in H1299 cell lysate.

Western Blotting Analysis: A 1:5,000 dilution of this antibody detected KRAB family zinc finger proteins in 10 µg of H1299 cell lysate.

Concentration: Please refer to lot specific datasheet.

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.