biological source
rabbit
Quality Level
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 μL
technique(s)
immunofluorescence: suitable
western blot: suitable
isotype
IgG
epitope sequence
N-terminal
Protein ID accession no.
UniProt accession no.
shipped in
2-8°C
target post-translational modification
mutation (Lys4Met)
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
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.
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Storage Class Code
12 - Non Combustible Liquids
WGK
WGK 1
Flash Point(F)
Not applicable
Flash Point(C)
Not applicable
Certificates of Analysis (COA)
Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.
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