Anti-Human p-Histone H3-Ser10 (3H10) ColorWheel^® Dye-Ready mAb

ColorWheel^® technology is a novel and proprietary method of creating your own antibody and dye combinations for use in flow cytometry. By incubating any ColorWheel^® antibody with any ColorWheel^® dye, researchers can quickly and simply produce primary conjugated antibodies for use in single color or multicolor flow cytometry analysis. Each ColorWheel^® antibody and dye is lyophilized for long-term storage, allowing for simplicity and flexibility without compromise. ColorWheel^® technology requires both ColorWheel^® antibody and ColorWheel^® dye for Flow Cytometry application.

Histone H3.1t (UniProt: Q16695; also known as H3/t, H3t, H3/g) is encoded by the HIST3H3 (also known as H3FT) gene (Gene ID: 8290) in human. Histones are highly conserved basic nuclear proteins that are responsible for the nucleosome structure of chromatin in eukaryotes. They play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. Two molecules of each of the four core histones (H2A, H2B, H3, and H4) form an octamer, around which DNA is wrapped in repeating units, called nucleosomes, which limits DNA accessibility to the cellular machineries that require DNA as a template. Histone H3 features a main globular domain and a long N-terminal tail, which protrudes from the globular nucleosome core and can undergo several different types of epigenetic modifications that influence cellular processes. The amino-terminal tails of histone proteins are subject to posttranslational modifications, including phosphorylation, acetylation, and methylation, which recruit downstream regulatory factors, influence chromatin structure, and are critical determinants of transcription. Histone H3 can undergo acetylation on several lysine residues in the histone tail by histone acetyltransferases. Acetylation is generally associated with transcriptional activity and methylation of lysine and arginine residues can either activate or repress depending on the residue modified. Trimethylation of histone H3 on Lys9 (H3K9me3) is one of the most studied epigenetic marks. Histone phosphorylation at serine 10 (with initiator methionine removed) by Aurora kinase B is crucial for chromosome condensation and cell-cycle progression during mitosis and meiosis. In addition, phosphorylation at this residue by RPS6KA4 and RPS6KA5 is essential 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. Phosphorylation of serine 10 is reported to inhibits Chk1 and PKC-mediated modification of Thr11 and Thr6 residues, the respective primary substrate sites of these kinases. This ColorWheel^® Dye-Ready monoclonal antibody was developed using our Cat. No. 05-806; Anti-phospho-Histone H3 (Ser10), clone 3H10. (Ref.: Liokatis, S., et al. (2012). Nat. Struct. Mol. Biol. 19(8); 819-823; Wiencke, JK., et al. (2008). Oncogene. 27(17); 2412-2421; Tamaru, H., et al. (2003). Nat. Genet. 34(1); 75-79).

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A linear peptide corresponding a sequence surrounding phosphoserine 10 of human Histone H3.

Quality Control Testing

Evaluated by Flow Cytometry in HeLa cells.

Flow Cytometry Analysis: One million HeLa cells, either untreated or treated with nocodazole (10 µM; 24 h), fixed and permeabilized, were stained with 5 µL of a 1:1 mixture of Cat. No. CWA-1173, Anti-Human p-Histone H3-Ser10 (3H10) ColorWheel^® Dye-Ready mAb and Cat. No. CWDS-PE ColorWheel^® Antibody-Ready Phycoerythrin (PE) Dye or an equivalent amount of Cat. No. CWA-1013 Mouse IgG1 isotype control and Cat. No. CWDS-PE ColorWheel^® Antibody-Ready Phycoerythrin (PE) Dye.

Tested Applications

Flow Cytometry Analysis: One million HeLa cells, either untreated or treated with nocodazole (10 µM; 24 h), fixed and permeabilized, were stained with 5 µL of a 1:1 mixture of Cat. No. CWA-1173, Anti-Human p-Histone H3-Ser10 (3H10) ColorWheel^® Dye-Ready mAb and Cat. No. CWDS-APC ColorWheel^® Antibody-Ready Allophycocyanin (APC) Dye or the equivalent amount of Cat. No. CWA-1013 Mouse IgG1 isotype control and Cat. No. CWDS-APC ColorWheel^® Antibody-Ready Allophycocyanin (APC) Dye.

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

Clone 3H10 is a mouse monoclonal antibody that detects Histone H3 phosphorylated on serine 10.

Lyophilized from PBS containing D-Mannitol and Sucrose, normal appearance is a dried pellet. Reconstituted antibody solution is stable and functional as assessed by functional testing. Contains no biocide or preservatives, such as azide.

1.0 mg/mL after reconstitution at 25 µL in PBS. 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 PBS. Protect from light.

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

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.