ZRB2924

Anti-p-Histone H3.3 (S31) Antibody, clone 2I12 ZooMAb^® Rabbit Monoclonal

recombinant, expressed in HEK 293 cells

• Synonym(s): H3.3A, H3F3, H3F3A, H3.3B, H3F3B, PP781
• UNSPSC Code: 12352203

Properties

• biological source: rabbit
• Quality Level: 200
• recombinant: expressed in HEK 293 cells
• conjugate: unconjugated
• antibody form: purified antibody
• antibody product type: primary antibodies
• clone: 2I12-H1, recombinant monoclonal
• description: recombinant, expressed in HEK 293 cells
• product line: ZooMAb^® learn more
• form: lyophilized
• mol wt: calculated mol wt 15 kDa; observed mol wt ~15 kDa
• purified by: using Protein A
• species reactivity: human
• species reactivity (predicted by homology): mouse, rat
• packaging: antibody small pack of 25
• greener alternative product characteristics: Waste Prevention; Designing Safer Chemicals; Design for Energy Efficiency; Learn more about the Principles of Green Chemistry.
• enhanced validation: recombinant expression; Learn more about Antibody Enhanced Validation
• sustainability: Greener Alternative Product
• technique(s): immunocytochemistry: suitable; immunohistochemistry: suitable; inhibition assay: suitable; western blot: suitable
• isotype: IgG
• epitope sequence: N-terminal half
• Protein ID accession no.: NP_001365972 ( But uniprot shows NP_005315, NP_002098)
• UniProt accession no.: P84243
• greener alternative category: Aligned
• shipped in: ambient
• storage temp.: 2-8°C

Description

General description

We are committed to bringing you greener alternative products, which adhere to one or more of the 12 Principles of Green Chemistry. This antibody is preservative-free, produced without the harm or sacrifice of animals and exceptionally stable to allow for ambient shipping and storage if needed, and thus aligns with "Waste Prevention", "Designing Safer Chemicals" and "Design for Energy Efficiency". Click here for more information.

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.

Specificity

Clone 2I12-H1 is a ZooMAb^® rabbit recombinant monoclonal antibody that detects phospho-Histone H3.3 (Ser31). It targets an epitope within 11 amino acids from the N-terminal half.

Immunogen

KLH-conjugated linear peptide corresponding to 11 amino acids from the N-terminal half of human phospho-Histone H3.3 (Ser31).

Application

Quality Control Testing

Evaluated by Western Blotting in cell lysate.

Western Blotting Analysis: A 1:500 dilution of this antibody detected phospho-Histone H3.3 (Ser31) in acid extract of HeLa cells treated with Colcemid (50 ng/mL; overnight).

Tested Applications

Peptide Inhibition Assay: Target band detection in lysate from acid extract of HeLa cells treated with colcemid (50 ng/mL; overnight) was prevented by pre-blocking of a representative lot with the immunogen phosphopeptide, but not the corresponding non-phosphopeptide.

Immunocytochemistry Analysis: A 1:100 dilution from a representative lot detected phospho-Histone H3.3 (Ser31) in HeLa cells.

Immunohistochemistry (Paraffin) Analysis: A 1:100 dilution from a representative lot detected phospho-Histone H3.3 (Ser31) in human colon tissue sections.

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

Target description

Histone H3.3 (UniProt: P84243) is encoded by the H3-3A; H3-3B (also known as H3.3A, H3F3, H3F3A, PP781, H3.3B, H3F3B) gene (Gene ID: 3020, 3021) in human. Histone H3 is one of the five main histone proteins involved in the structure of chromatin in eukaryotic cells. 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 H3 variants (H3.1, H3.2 and H3.3) have also been implicated in the epigenetic memory of cellular state. The Ser31 residue of H3.3 (H3.3S31) is phosphorylated by the Chk1 kinase that suggests a role in transcriptional gene activation. H3.3S31 phosphorylation influences chromosome segregation during mitosis and regulates heterochromatin accessibility at telomeres during replication. H3.3S31ph is observed in late prometaphase and metaphase, but not in anaphase. This ZooMAb^® recombinant monoclonal antibody, generated by our proprietary technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. (Ref.: Martire, S., et al., (2019), Nat Genet., 51:6, 941 946.; Hake, S.B., et al., (2005), PNAS, 102:18, 6344-6349)

Physical form

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.

Reconstitution

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.

Storage and Stability

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.

Legal Information

ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany

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.