Bromodeoxyuridine (BrDU) Antikörper

Antigen: Bromodeoxyuridine (BrDU)

Klonalität: Polyklonal

Wirt: Schaf

Reaktivität: Human

Applikation: Immunhistochemie (Gefrierschnitte) (IHC (fro)), Enzyme Immunoassay (EIA), Immunfluoreszenz (IF), Immunpräzipitation (IP), Immunhistochemie (Paraffinschnitte) (IHC (p)), Western Blot (WB)

Produktnummer: ABIN153035

Produktbeschreibung

Immunogen: Bromodeoxyuridine coupled to keyhole limpet hemocyanin (KLH).

Beschreibung: To measure DNA synthesis or cell proliferation, 5-bromo2’-deoxy-uridine (BrdU) can beincorporated into DNA in place of thymindine. Cells, which have incorporated BrdU intoDNA, can be quickly detected using a monoclonal antibody against BrdU. The binding ofthe antibody is achieved by denaturation of the DNA. This is usually obtained byexposing the cells to acid, or heat. Alternate Names: anti-Bromodeoxyuridine antibody, anti-BUdr antibody.

Spezifität: Species Reactivity: All.

Anwendungen

Applikationshinweise: Fixation in cold methanol for 30 minutes followed by immersion in 7x10-3 M NaOH for10-15 seconds allows BrdU staining with the simultaneous detection of nuclearcytoplasmic and membrane assigns as well as preservation of morphological detail.The unconjugated antibody was tested using immunoprecipitation against 5-MethylCytosine (5-MeC) and bromodeoxyuridine (BrdU) or control (no antigen) and assayedby A-405 spectrophotometry. At a concentration of 25 ug/mL, this product demonstrates8 fold higher reactivity against BrdU than 5-MeC. Product titers out to 50 ng/mL in thepurified form.For best results on unconjugated antibody, use the product at a concentration of 25 to100 ug/mL. Nearly complete immunoprecipitation was obtained at concentrations of 100to 500ug/mL. The product has also been tested for utility for staining ofbromodeoxyuridine incorporated into DNA of replicating cells. When utilized at a purifiedconcentration of 10 ug/mL, the product is comparable to commercially availablemonoclonal antibodies commonly used for the same purpose.A key issue is that it is necessary to denature the DNA so as to expose the base to theantibodies. One protocol is: Sachiko Matsu uraa and Kazuo SuzukiaImmunohistochemical Analysis of DNA Synthesis During Chronic Stimulation withIsoproterenol in Mouse Submandibular Gland . Journal of Histochemistry andCytochemistry, Vol. 45, 1137-1146, Immunohistochemistry DNA-synthesizing cells weredetected by peroxidase-anti-peroxidase (PAP) immunostaining with anti-BrdUantibodies by the method of Harms et al. 1986 , which was slightly modified as follows:Sections were treated with 2 N HCl at 60C for 15 min and washed in running water for15 min. After dehydration in an ethanol series, the sections were etched with xylene (15min, twice). Nuclear proteins, which mask incorporated BrdU, were digested with0.025% protease (Type V, Sigma) in PBS at 37C for 10 min, and the sections were thenwashed with cold PBS three times for 5 min.

Konzentration: 1.93 mg/ml

Buffer: 0.15M PBS. pH 7.5. Preservative: No preservatives.

Lagerung: Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.

Forschungsgebiet: Proliferationsmarker

Beschränkungen: Nur für Forschungszwecke einsetzbar

Bilder

anti-Bromodeoxyuridine (BrDU) antibody

Publikationen

Publikationen: Campana, Coustan-Smith, Janossy: "Double and triple staining methods for studying the proliferative activity of human B and T lymphoid cells." in: Journal of immunological methods, Vol. 107, Issue 1, pp. 79-88, 1988 (PubMed).

Morris, Solomon: "BRCA1 : BARD1 induces the formation of conjugated ubiquitin structures, dependent on K6 of ubiquitin, in cells during DNA replication and repair." in: Human molecular genetics, Vol. 13, Issue 8, pp. 807-17, 2004 (PubMed).

Cooper, Isacson: "Intrastriatal transforming growth factor alpha delivery to a model of Parkinson's disease induces proliferation and migration of endogenous adult neural progenitor cells without differentiation into dopaminergic neurons." in: The Journal of neuroscience : the official journal of the Society for Neuroscience, Vol. 24, Issue 41, pp. 8924-31, 2004 (PubMed).