ANTI-ZNF189 25 Y56 (C15-1317-117)

General description

Histone H3.3 (UniProt: P84243; also known as H3.3) is encoded by the H3F3A (also known as H3.3A, H3F3, PP781, H3F3B, H3.3B) gene (Gene ID: 3020; 3021) in human. Histone H3.3 is a highly conserved variant form of Histone H3, which replaces conventional H3 in a wide range of nucleosomes in active genes. Histone H3 has two main variants, H3.1 and H3.3, which show different genomic localization patterns in animals. The H3.1 and H3.3 complexes also possess distinct histone chaperones, CAF-1 and HIRA, which play important role in mediating DNA-synthesis-dependent and -independent nucleosome assembly. Histone H3.1 serves as the canonical histone, which is incorporated during DNA replication, whereas H3.3 acts as the replacement histone that can be incorporated outside of S-phase during chromatin-disrupting processes like transcription. Histone H 3.3 constitutes the predominant form of histone H3 in non-dividing cells and is incorporated into chromatin independently of DNA synthesis. It is associated with actively expressed genes in both animals and plants. It is predominantly enriched near transcription end sites (TES) of genes and positively associated with transcription. Histone H3 contains a main globular domain and a long N-terminal tail and is involved with the structure of the nucleosomes of the ′beads on a string′ structure. The N-terminal tail of histone H3 protrudes from the globular nucleosome core and can undergo several different types of epigenetic modifications that influence cellular processes. These modifications include the covalent attachment of methyl or acetyl groups to lysine and arginine amino acids and the phosphorylation of serine or threonine. Mutations in Histone H3.3 have been implicated in a high proportion of malignant pediatric brain cancers. K4M mutation is reported to inhibit H3K4 methylation and prevent enhancer activation in adipogenesis by destabilizing MLL4/MLL4 proteins, but does not affect MLL1, MLL2, SETA1, or SET1B. Expression of K4M in lineage-specific precursor cells is shown to deplete H3K4 methylation and impair adipose tissue and muscle development. (Ref.: Jang, Y., et al. (2019). Nucleic Acids Res. 47(2); 607-620; Tagami, H., et al. (2004). Cell, 116(1); 51-61).

Specificity

This rabbit polyclonal antibody detected K4M and K4I mutant forms of Histone H3.

Immunogen

KLH-conjugated linear peptide corresponding to the first 9 amino acids (without first methionine) from the N-terminal region of Histone H3 with K4M mutation.

Application

Quality Control Testing

Evaluated by Western Blotting in HEK293T cells transfected with K4M mutant.

Western Blotting Analysis: A 1:1,000 dilution of this antibody detected Histone H3.3 in HEK293T cells transfected with K4M mutation.

Tested Applications

Immunofluorescence Analysis: A representative lot detected Histone H3.3 in HEK293T cells transfected with K4M mutant (Courtesy of Dave Allis Lab @ Rockefeller University).

Western Blotting Analysis: A 1:2,000 dilution from a representative lot detected Histone H3.3 in HEK293T cells transfected with K4M mutant (Courtesy of Dave Allis Lab @ Rockefeller University).

Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user

Anti-Histone H3 K4M mutant, Cat. No. ABE2927, is a rabbit polyclonal antibody that detects Histone H3 with K4M mutation and is tested for use in Immunofluorescence and Western Blotting.

Physical form

Purified rabbit polyclonal antibody in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

Storage and Stability

Recommend storage at +2°C to +8°C. For long term storage antibodies can be kept at -20°C. Avoid repeated freeze-thaws.

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Disclaimer

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.

biological source: rabbit. Quality Level: 200. conjugate: unconjugated. antibody form: purified antibody. antibody product type: primary antibodies. clone: polyclonal. mol wt: calculated mol wt 15.33 . kDa, observed mol wt ~18 . kDa. purified by: affinity chromatography. species reactivity: human. packaging: antibody small pack of 100 . μ. L. technique(s): immunofluorescence: suitable, western blot: suitable. isotype: IgG. epitope sequence: N-terminal. Protein ID accession no.: NP_001365972. UniProt accession no.: P84243. shipped in: 2-8°C. target post-translational modification: mutation (Lys4Met). Storage Class Code: 12 - Non Combustible Liquids. WGK: WGK 1. Flash Point(F): Not applicable. Flash Point(C): Not applicable.