Histone 3 anticorps (pSer28) (Alexa Fluor 647)

N° du produit ABIN1177066

Cet anticorps Rat Monoclonal détecte spécifiquement Histone 3 dans ICS. Il présente une réactivité avec des échantillons de Humain, Souris, Rat, Boeuf (Vache), Hamster et Blow Fly. Il a été cité dans 9+ publications.

Aperçu rapide pour Histone 3 anticorps (pSer28) (Alexa Fluor 647) (ABIN1177066)

Antigène: Histone 3 (H3) (Histone H3 (H3))

Reactivité: Humain, Souris, Rat, Boeuf (Vache), Hamster, Blow Fly

Hôte: Rat

Clonalité: Monoclonal

Conjugué: Cet anticorp Histone 3 est conjugé à/à la Alexa Fluor 647

Application: Intracellular Staining (ICS)

Clone: HTA28

Détail du produit anti-Histone 3 anticorps (Alexa Fluor 647)

Épitope: pSer28

Marque: BD Phosflow™

Purification: The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Immunogène: Phosphorylated Human Histone H3 Peptide

Isotype: IgG2a kappa

Information d'application

Volume d'échantillon: 20 μL

Protocole: 1. Wash cell suspension twice with 1X PBS.
2. Fix the cells by adding ice-cold 70% ethanol drop-wise while vortexing the cell suspension, then storing them for at least 4 hours at -20°C in the 70% ethanol.
3. Aliquot ~1 million fixed cells per tube for staining. Wash them twice with 1X PBS, then once with stain buffer.
4. Stain the cells with 20 myl Alexa Fluor® 647 Rat anti-Histone H3 (pS28) in 80 myl stain buffer for 20 minutes at room temperature, then wash them with stain buffer.
5. For optimum cell cycle analysis, the cells should be treated with RNAse before staining with propidium iodide:
a) Treat the stained cells with 50 myg RNAse A (Sigma R5500) in 50 myl 1X PBS for 30 minutes at 37°C. Without washing, stain DNA by adding 5 myg Propidium Iodide (Sigma P4170) in 450 myl staining buffer for at least 10 minutes at room temperature.
b) Stain the cells with 0.5 ml PI/RNase Staining Buffer (Cat. No. 550825) for 15 minutes at room temperature.
6. The cells are now ready for flow cytometric analysis.

Restrictions: For Research Use only

Stockage

Format: Liquid

Buffer: Aqueous buffered solution containing BSA and ≤0.09 % sodium azide.

Agent conservateur: Sodium azide

Précaution d'utilisation: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Stock: 4 °C

Stockage commentaire: Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The antibody was conjugated to Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.

Références pour anticorps anti-Histone 3 (Alexa Fluor 647) (ABIN1177066)

Dyson, Thomson, Inagaki, Goto, Arthur, Nightingale, Iborra, Mahadevan: "MAP kinase-mediated phosphorylation of distinct pools of histone H3 at S10 or S28 via mitogen- and stress-activated kinase 1/2." dans: Journal of cell science, Vol. 118, Issue Pt 10, pp. 2247-59, (2005) (PubMed).

Choi, Choi, Cho, Zhu, Bode, Dong: "Phosphorylation of Ser28 in histone H3 mediated by mixed lineage kinase-like mitogen-activated protein triple kinase alpha." dans: The Journal of biological chemistry, Vol. 280, Issue 14, pp. 13545-53, (2005) (PubMed).

Nowak, Corces: "Phosphorylation of histone H3: a balancing act between chromosome condensation and transcriptional activation." dans: Trends in genetics : TIG, Vol. 20, Issue 4, pp. 214-20, (2004) (PubMed).

Hirata, Inada, Tsukamoto, Sakai, Mizoshita, Yanai, Masegi, Goto, Inagaki, Tatematsu: "Characterization of a monoclonal antibody, HTA28, recognizing a histone H3 phosphorylation site as a useful marker of M-phase cells." dans: The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society, Vol. 52, Issue 11, pp. 1503-9, (2004) (PubMed).

Carmena, Earnshaw: "The cellular geography of aurora kinases." dans: Nature reviews. Molecular cell biology, Vol. 4, Issue 11, pp. 842-54, (2003) (PubMed).

Furukawa, Sugiyama, Osouda, Goto, Inagaki, Horigome, Omata, McConnell, Fisher, Nishida: "Barrier-to-autointegration factor plays crucial roles in cell cycle progression and nuclear organization in Drosophila." dans: Journal of cell science, Vol. 116, Issue Pt 18, pp. 3811-23, (2003) (PubMed).

Pascreau, Arlot-Bonnemains, Prigent: "Phosphorylation of histone and histone-like proteins by aurora kinases during mitosis." dans: Progress in cell cycle research, Vol. 5, pp. 369-74, (2003) (PubMed).

Cheung, Allis, Sassone-Corsi: "Signaling to chromatin through histone modifications." dans: Cell, Vol. 103, Issue 2, pp. 263-71, (2000) (PubMed).

Goto, Tomono, Ajiro, Kosako, Fujita, Sakurai, Okawa, Iwamatsu, Okigaki, Takahashi, Inagaki: "Identification of a novel phosphorylation site on histone H3 coupled with mitotic chromosome condensation." dans: The Journal of biological chemistry, Vol. 274, Issue 36, pp. 25543-9, (1999) (PubMed).

Détails sur Histone 3

Antigène: Histone 3 (H3) (Histone H3 (H3))

Autre désignation: Histone H3

Sujet: Histones are highly basic proteins that complex with DNA to form chromatin. Histone H3 is a ~15-kDa protein that is phosphorylated at serine 28 (S28), S10, and/or threonine 11 during mammalian cell mitosis and meiosis. The phosphorylation sites are located in the N-terminal tail, a region that is outside of the chromatin fiber and is thus accessible for interactions with agents that may regulate chromatin or specific gene activities. The phosphorylation states of the two serine sites during the cell cycle are highly regulated by Aurora B kinase and a PP1 phosphatase: S10 is in the phosphorylated state from late G2 phase to anaphase, while S28 is phosphorylated from prophase to anaphase. Furthermore, phosphorylation of histone H3 S28 may be mediated by other kinases in response to external stimuli. Evidence suggests that histone phosphorylation is involved in the regulation of chromosome condensation, cell division, and gene transcription. The HTA28 monoclonal antibody reacts with histone H3 phosphorylated at S28 in its N-terminal tail. It does not recognize the unphosphorylated protein.