500095 | OxyDNA Assay Kit, Fluorometric

500095
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      Prix & Disponibilité

      RéférenceDisponibilité Conditionnement Qté Prix Quantité
      500095-1KIT
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      Disponibilité limitée Disponibilité limitée
      Interrompu(e)
      Disponible en quantités limitées
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          1 kit
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          Description
          Aperçu Assay kit used to detect oxidative damage to DNA in vitro. Based on the direct binding of a fluorescent probe to the DNA adduct 8-oxoguanine, a major oxidation product and an important indicator of free radical-induced DNA damage and oxidative stress. The FITC-conjugate binds to 8-oxoguanine in damaged cells and fluorescence is monitored using flow cytometery (excitation 495 nm, emission 515 nm).
          Référence 500095
          Famille de marques Calbiochem®
          Références bibliographiques
          Références bibliographiques Silva, P.F.N. et al. 2007. Theriogenology 67, 609.
          Babbar, N. and Casero, Jr., R.A. 2006. Cancer Res. 66, 11125.
          Lai, K.N., et al. 2006. Nephrol. Dial. Transplant. 21, 1188.
          Sampson, M.J. et al. 2006. Diabetes Care 29, 283.
          Maniscalco, W.M., et al. 2005. Pediatric Res. 58, 594.
          Osella, D., et al. 2005. Inorganica Chimica Acta 358, 1993.
          Adamek B et al. 2004. Polish Journal of Environmental Studies 13 Suppl. II, 29.
          Roper, J.M., et al. 2004. Am. J. Physiol. Lung Cell Mol. Physiol. 286, L1045.
          Ogawa, Y. et al. 2003. International Journal of Molecular Medicine 11, 27.
          Ogawa, Y. et al. 2003. International Journal of Molecular Medicine 11, 149.
          Sparrow J.R. et al. 2003. Investigative Ophthalmology and Visual Science (14)5, 2245.
          Tao Peng, H-M. et al. 2003. World J Gastroenterol 9, 2186.
          Riemschneider, S., et al. 2002. Acta Derm. Venereol. 82, 325.
          de Zwart, L.L., et al. 1999. Free Radic. Biol. Med. 26, 202.
          Tsurudome, Y., et al. 1999. Carcinogenesis 20, 1573.
          Inoue, M., et al. 1998. Jpn. J. Cancer Res. 89, 691.
          Kasai, H. 1997. Mutat. Res. 387, 147.
          Cooke, C., et al. 1996. Toxicol. Ecotoxicol. News 3, 101.



          Selected Citations
          Roper, J. M., et al. 2004. Am. J. Physiol. Lung Cell Mol. Physiol. 286, L1045.
          Babbar, N., and Casero, R. A., Jr., 2006. Cancer Res. 66, 23.


          Informations produit
          Méthode de détection Flow cytometry or fluorescence microscopy
          Forme 50 Tests
          Contenu du kit Wash Concentrate, Binding Protein-FITC Conjugate, and a user protocol.
          Applications
          Références d'applications Fluorescent Microscopy Lai, K.N., et al. 2006. Nephrol. Dial. Transplant. 21, 1188. Maniscalco, W.M., et al. 2005. Pediatric Res. 58, 594. Roper, J.M., et al. 2004. Am. J. Physiol. Lung Cell Mol. Physiol. 286, L1045. Other Applications Babbar, N. and Casero, Jr., R.A. 2006. Cancer Res. 66, 11125. Osella, D., et al. 2005. Inorganica Chimica Acta 358, 1993. Riemschneider, S., et al. 2002. Acta Derm. Venereol. 82, 325.
          Informations biologiques
          Temps de l'essai 4 h
          Type d'échantillon Cells
          Informations physico-chimiques
          Dimensions
          Informations sur les matériaux
          Informations toxicologiques
          Informations de sécurité selon le SGH
          Informations sur la sécurité
          Notifications sur l'utilisation du produit
          Utilisation prévue The Calbiochem® OxyDNA Kit is an in vitro fluorescent protein binding method for the detection of oxidative damage to DNA in fixed permeabilized cells using flow cytometry or fluorescence microscopy. Material of both human and animal origin can be tested.
          Informations relatives au stockage et à l'expédition
          Code Expé Ambient Temperature Only
          Toxicité Multiple Toxicity Values, refer to MSDS
          Stockage +2°C to +8°C
          Conditions de stockage Upon arrival store the entire contents of the kit at 4°C. Note: FITC conjugate must be stored in the dark.
          Ne pas congeler No
          Informations sur l'emballage
          Informations sur le transport
          Information complémentaire
          Contenu du kit Wash Concentrate, Binding Protein-FITC Conjugate, and a user protocol.
          Caractéristiques

          Documentation

          FDS

          Langues
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          Certificats d'analyse

          TitreNuméro de lot
          500095

          Références bibliographiques

          Aperçu de la référence bibliographique
          Silva, P.F.N. et al. 2007. Theriogenology 67, 609.
          Babbar, N. and Casero, Jr., R.A. 2006. Cancer Res. 66, 11125.
          Lai, K.N., et al. 2006. Nephrol. Dial. Transplant. 21, 1188.
          Sampson, M.J. et al. 2006. Diabetes Care 29, 283.
          Maniscalco, W.M., et al. 2005. Pediatric Res. 58, 594.
          Osella, D., et al. 2005. Inorganica Chimica Acta 358, 1993.
          Adamek B et al. 2004. Polish Journal of Environmental Studies 13 Suppl. II, 29.
          Roper, J.M., et al. 2004. Am. J. Physiol. Lung Cell Mol. Physiol. 286, L1045.
          Ogawa, Y. et al. 2003. International Journal of Molecular Medicine 11, 27.
          Ogawa, Y. et al. 2003. International Journal of Molecular Medicine 11, 149.
          Sparrow J.R. et al. 2003. Investigative Ophthalmology and Visual Science (14)5, 2245.
          Tao Peng, H-M. et al. 2003. World J Gastroenterol 9, 2186.
          Riemschneider, S., et al. 2002. Acta Derm. Venereol. 82, 325.
          de Zwart, L.L., et al. 1999. Free Radic. Biol. Med. 26, 202.
          Tsurudome, Y., et al. 1999. Carcinogenesis 20, 1573.
          Inoue, M., et al. 1998. Jpn. J. Cancer Res. 89, 691.
          Kasai, H. 1997. Mutat. Res. 387, 147.
          Cooke, C., et al. 1996. Toxicol. Ecotoxicol. News 3, 101.



          Selected Citations
          Roper, J. M., et al. 2004. Am. J. Physiol. Lung Cell Mol. Physiol. 286, L1045.
          Babbar, N., and Casero, R. A., Jr., 2006. Cancer Res. 66, 23.


          Protocole Utilisateur

          Révision 23-September-2014 JSW
          Forme 50 Tests
          Méthode de détection Flow cytometry or fluorescence microscopy
          Stockage Upon arrival store the entire contents of the kit at 4°C. Note: FITC conjugate must be stored in the dark.
          Utilisation prévue The Calbiochem® OxyDNA Kit is an in vitro fluorescent protein binding method for the detection of oxidative damage to DNA in fixed permeabilized cells using flow cytometry or fluorescence microscopy. Material of both human and animal origin can be tested.
          Contexte Oxidative injury to macromolecules is indicated in a wide range of pathological conditions. Damage is mediated via free radicals that can be created by a range of agents, e.g. xenobiotics, environmental toxins, smoking, radiation, ischaemia-reperfusion injury oxidising agents and normal or disturbed metabolic activity. These free radicals may react with DNA causing reversible and irreversible damage. This can lead to mutation, carcinogenesis, teratogenesis or cell death. The probe in the OxyDNA kit is specific for 8-oxoguanine. 8-oxoguanine (as part of the oxidized nucleotide 8-oxyguanosine) is formed during free radical damage to DNA and is a sensitive and specific indicator of oxidative DNA damage. 8-oxoguanine is a particularly important biomarker of oxidative DNA damage as it is formed in relatively large quantities and 8-oxoguanine formation can lead to mutations, such as the substitution of thymine for guanine and cytosine for adenine. Previously, 8-oxoguanine was difficult to detect, requiring HPLC analysis, however by utilizing a binding protein with high avidity and specificity for 8-oxoguanine, the OxyDNA kit provides a simple, convenient, sensitive fluorescence method for detecting for oxidative DNA damage.
          Principes de l'essai The Calbiochem® OxyDNA Kit utilizes a direct fluorescent protein binding technique. After cells have been fixed and permeabilized, the FITC-Conjugate is added and binds to the 8-oxoguanine moiety present in the 8-oxoguanosine of oxidized DNA. The presence of oxidized DNA is indicated by a green/yellow fluorescence that can be read using flow cytometry or fluorescence microscopy.
          Matériel fourni • Wash Concentrate (Kit component No. KP10401-55ML): 1 bottle, 55 ml, 25X Tris-buffered Saline/TWEEN® 20 Detergent (TBST), containing ProClin950 concentrate
          • FITC-Conjugate Concentrate (Kit Component No. KP10402-500UL): 1 vial, 0.5 ml Binding Protein conjugated to Fluorescein, containing ProClin950 and Bronidox L.
          Matériel requis, mais non fourni Micropipettes: 20 µl to 100 µl, 200 µl to 1000 µl
          Test tubes
          1 L beaker
          Graduated cylinder
          Reagents for fixing and permeabilising cells
          Deionized/Distilled water
          37°C water bath
          Flow cytometer with appropriate filter combination for FITC (excitation filter 495 nm, barrier filter 515 nm)
          Précautions et recommandations The OxyDNA kit contains a chemical known to the State of California to cause birth defects or other reproductive harm (California Prop 65: thimerosal).
          Dispose of all specimens in accordance with good laboratory practice.
          Wear protective clothing, disposable latex gloves and eye protection while handling specimens and performing the assay. Wash hands thoroughly when finished.
          Do not pipette materials by mouth and never eat or drink at the laboratory workbench.
          Do not mix or substitute reagents from different kit lot numbers.
          Care must be taken not to contaminate components and always use fresh pipette tips for each sample and component.
          Do not use reagents that are cloudy or that have precipitated out of solution. High quality distilled or deionised water is required for the Wash Solution. The use of poor quality or contaminated water may lead to background colour in the assay.
          Allow all reagents to come to room temperature (20-25°C) and mix well prior to use.
          Avoid leaving reagents in direct sunlight and/or above 4°C for extended periods.
          Always use clean, preferably disposable, glassware for all reagent preparation.
          Préparation Live cells should be shielded from high oxygen tensions (e.g. room air) and unnecessary mechanical stress (e.g. mixing and washing) as these can lead to increased oxidative DNA damage and high levels of fluorescence in untreated/control samples. DNA is more stable in fixed cells.
          Préparation des réactifs • Wash Solution: Dilute the Wash Concentrate 1:25 with diH2O. For example, to prepare 250 ml Wash Solution add 10 ml of Wash Concentrate to 240 ml diH2O. Prepare only the volume of Wash Solution required for the assay. • 1X FITC-Conjugate: Dilute the FITC-Conjugate 1:10 with Wash Solution. For example, to prepare 1 ml 1X FITC-Conjugate add 100 µl FITC-Conjugate to 900 µl Wash Solution. To dispense conjugate, remove cap and pipette directly from the bottle. Keep FITC-Conjugate solutions in the dark when not in use. These dilutions of FITC-Conjugate have been used successfully for a number of cell lines e.g. Hep G2, IMR-32 and CHO, sperm and lymphocytes. In some experimental situations, it may be necessary to optimize the dilution of the conjugate used. This will be dependent on the particular cell type used and the experimental conditions used to induce oxidative damage. If the incorrect concentration of conjugate is used, high background staining and non-specific binding may occur.
          Protocole détaillé Assay Protocol for Flow Cytometry Assay

          Oxidative DNA damage can be found in most cell types and can be caused by many different mechanisms. As a result, the protocol outlined below should only be regarded as a guideline and the procedures may need to be modified according to the experimental model or cell type being studied.

          1. Fix and permeabilize the cells to be tested.
          2. Wash with Wash Solution.
          3. Add 100 µl 1X FITC-Conjugate to the cell pellet and incubate in the dark for 60 min at room temperature.
          4. Wash with Wash Solution.
          5. Read fluorescence using a flow cytometer at an excitation wavelength of 495 nm and barrier filter of 515 nm.
          Spécificité The Calbiochem® OxyDNA Kit is specific for the 8-oxoguanine moiety of 8-oxoguanosine present in oxidized DNA and shows no measurable cross reactivity with unoxidized guanine / guanosine or other unoxidized nucleotides.
          Références d'applications Fluorescent Microscopy Lai, K.N., et al. 2006. Nephrol. Dial. Transplant. 21, 1188. Maniscalco, W.M., et al. 2005. Pediatric Res. 58, 594. Roper, J.M., et al. 2004. Am. J. Physiol. Lung Cell Mol. Physiol. 286, L1045. Other Applications Babbar, N. and Casero, Jr., R.A. 2006. Cancer Res. 66, 11125. Osella, D., et al. 2005. Inorganica Chimica Acta 358, 1993. Riemschneider, S., et al. 2002. Acta Derm. Venereol. 82, 325.
          Marques déposées Calbiochem® is a registered trademark of EMD Chemicals, Inc.
          Tween® is a registered trademark of ICI Americas, Inc.
          Interactive Pathways™ is a trademark of EMD Chemicals, Inc.

          Produits & Applications associés

          Produits associés classés par : Application Facette