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ABE419

Sigma-Aldrich

Anti-Histone H3.3 Antibody, K27M mutant

from rabbit, purified by affinity chromatography

Synonym(s):

Histone H3.1, Histone H3.3

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About This Item

UNSPSC Code:
12352203
eCl@ss:
32160702
NACRES:
NA.41

biological source

rabbit

Quality Level

100

antibody form

affinity isolated antibody

antibody product type

primary antibodies

clone

polyclonal

purified by

affinity chromatography

species reactivity

mouse, human

technique(s)

ChIP: suitable
immunohistochemistry: suitable
western blot: suitable

NCBI accession no.

NP_002098

UniProt accession no.

P84243

shipped in

wet ice

target post-translational modification

mutation (Lys27Met)

Gene Information

human ... H3F3B(3021)

General description

Histone H3.3 is one of the five main histone proteins involved in the structure of chromatin in eukaryotic cells. Featuring a main globular domain and a long N-terminal tail, H3.3 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. Histone variant H3.3 is typically enriched in active chromatin.

Specificity

This antibody recognizes Histone H3.3 with K27M mutation.

Immunogen

Epitope: Histsone H3 sequence surrounding K27M mutation
KLH-conjugated linear peptide corresponding to sequence near the N-terminus of human Histone H3.3 with K27M mutation.

Application

Anti-Histone H3.3 Antibody, K27M mutant, is validated for use in western blotting (WB) & Chromatin immunoprecipitation (ChIP).
Research Category
Epigenetics & Nuclear Function
Research Sub Category
Histones
Western Blotting Analysis: A representative lot detected histone H3.30 K27M mutant in diffuse intrinsic pontice Glioma tissue lysates expressing Histone H3.3 K27M mutant. (Lewis, P. W., et al. (2013). Science. 340(6134):857-861.

Chromatin Immunoprecipitation: A representative lot co-precipitated chromatin fragments containing the promoter regions of ACTA and CCT8 genes from HEK293T transfectants expressing FLAG-HA-tagged histone H3.3 with K27M mutation (Peter W. Lewis and David Allis, Laboratory of Chromatin Biology and Epigenetics, The Rockefeller University, New York, NY).

Quality

Evaluated by Western Blotting in MEF (H3.3) and MEF (H3.3 K27M-HA-Flag) transfected cell lysate.

Western Blotting Analysis: 0.2 µg/mL of this antibody detected FLAG-HA-tagged K27M mutant, but not wildtype, histone H3.3 in 3x10E5 cell equivalent of lysate from MEF transfectants.

Target description

~17 kDa observed

Physical form

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

Storage and Stability

Stable for 1 year at 2-8°C from date of receipt.

Analysis Note

Control
Lysates from MEF transfectants expressing K27M (positive) or wildtype (negative) FLAG-HA-tagged histone H3.3.

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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Abstracts from USCAP 2019: Neuropathology and Ophthalmic Pathology (1621-1666).
Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc, 32(Suppl 2), 1-39 (2019-03-20)
Specific detection of methionine 27 mutation in histone 3 variants (H3K27M) in fixed tissue from high-grade astrocytomas.
Bechet, D; Gielen, GG; Korshunov, A; Pfister, SM; Rousso, C; Faury, D; Fiset et al.
Acta neuropathologica null
Fausto J Rodriguez et al.
Brain pathology (Zurich, Switzerland), 29(1), 126-140 (2018-09-08)
Anaplasia may be identified in a subset of tumors with a presumed pilocytic astrocytoma (PA) component or piloid features, which may be associated with aggressive behavior, but the biologic basis of this change remains unclear. Fifty-seven resections from 36 patients
Sriram Venneti et al.
Acta neuropathologica, 128(5), 743-753 (2014-09-10)
Pediatric glioblastomas (GBM) are highly aggressive and lethal tumors. Recent sequencing studies have shown that ~30 % of pediatric GBM and ~80 % of diffuse intrinsic pontine gliomas show K27M mutations in the H3F3A gene, a variant encoding histone H3.3. H3F3A K27M
David A Solomon et al.
Brain pathology (Zurich, Switzerland), 26(5), 569-580 (2015-10-31)
Somatic mutations of the H3F3A and HIST1H3B genes encoding the histone H3 variants, H3.3 and H3.1, were recently identified in high-grade gliomas arising in the thalamus, pons and spinal cord of children and young adults. However, the complete range of
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