Anti-Histone H3.3 G34W Antibody, clone 2E20 ZooMAb^® Rabbit Monoclonal

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ZooMAb^® antibodies represent an entirely new generation of recombinant monoclonal antibodies. Each ZooMAb^® antibody is manufactured using our proprietary recombinant expression system, purified to homogeneity, and precisely dispensed to produce robust and highly reproducible lot-to-lot consistency. Only top-performing clones are released for use by researchers. Each antibody is validated for high specificity and affinity across multiple applications, including its most commonly used application. ZooMAb^® antibodies are reliably available and ready to ship when you need them.

Clone 2E20 is a ZooMAb^® rabbit recombinant monoclonal antibody that specifically detects Histone H3.3 with G34W mutation.

KLH-conjugated linear peptide corresponding to 8 amino acids surrounding the mutated G34W site from the N-terminal half of human Histone H3.3.

Quality Control Testing

Evaluated by Western Blotting in lysates from HEK293 cells expressing Histone H3.3 with G34W mutation.

Western Blotting Analysis (WB): A 1:10,000 dilution of this antibody detected Histone H3.3 G34W in lysate from HEK293 cells expressing this mutation, but not in wild-type HEK293 cells.

Tested Applications

Immunohistochemistry (Paraffin) Analysis: A 1:100 dilution from a representative lot detected Histone H3.3 G34W in Human bone giant cell tumor tissue sections.

Affinity Binding Assay: A representative lot of this antibody bound Histone H3.3 G34W peptide with a KD of 3.1 x 10-6 in an affinity binding assay.

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

Histone H3.3 (UniProt: P84243; also known as H3.3) is encoded by the H3-3A (also known as H3.3A, H3F3, H3F3A, PP781, H3-3B) gene (Gene ID: 3020) in human. Histone H3 has two main variants, H3.1 and H3.3, which show different genomic localization patterns in animals. Histone H3.1 serves as the canonical histone, which is incorporated during DNA replication. 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 H 3.3 constitutes the predominant form of histone H3 in non-dividing cells and is incorporated into chromatin independently of DNA synthesis. H3.3 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. For example, acetylation on Lys10 (H3K9ac; initial methionine removed) impairs methylation at Arg9 (H3R8me2s) and acetylation on Lys19 (H3K18ac) and Lys24 (H3K24ac) favors methylation at Arg18 (H3R17me). Phosphorylation at Ser11 (H3S10ph) by Aurora kinase B is crucial for chromosome condensation and cell-cycle progression during mitosis and meiosis. Phosphorylation at Ser11 by RPS6KA4 and RPS6KA5 is important during interphase because it enables the transcription of genes following external stimulation, like mitogens, stress, growth factors or UV irradiation and result in the activation of genes, such as c-Fos and c-Jun.  Mutations in Histone H3.3 have been implicated in a high proportion of malignant pediatric brain cancers. The mutant H3.3 histone disrupts epigenetic post-translational modifications near genes involved in cancer processes and in brain function. Glycine 34 to tryptophan (G34W) mutation in Histone 3.3 have been linked to the development of giant cell tumor of bone (GCTB) and also in a mosaic disorder characterized by pheochromocytomas and paragangliomas. This ZooMAb^® recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. (Ref.: Amary, F., et al. (2017). Am. J. Surg. Pathol. 41(8); 1059-1068).

Purified recombinant rabbit monoclonal antibody IgG, lyophilized in PBS with 5% Trehalose, normal appearance a coarse or translucent resin. The PBS/trehalose components in the ZooMAb formulation can have the appearance of a semi-solid (bead like gel) after lyophilization. This is a normal phenomenon. Please follow the recommended reconstitution procedure in the data sheet to dissolve the semi-solid, bead-like, gel-appearing material. The resulting antibody solution is completely stable and functional as proven by full functional testing. Contains no biocide or preservatives, such as azide, or any animal by-products. Larger pack sizes provided as multiples of 25 µL.

300 µg/mL after reconstitution at 25 µL per vial. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.

Recommend storage of lyophilized product at 2-8°C; Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial; Reconstitute each vial by adding 25 µL of filtered lab grade water or PBS; Reconstituted antibodies can be stored at 2-8°C, or -20°C for long term storage. Avoid repeated freeze-thaws.

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

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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.