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17-10228 LentiBrite™ Histone H2B-RFP Lentiviral Biosensor

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Key Applications: Transfection, Immunofluorescence, Immunocytochemistry
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17-10228
1 vial (minimum of 3 x 10E8 IFU/mL)  
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      Replacement Information

      Tableau de caractéristiques principal

      Key ApplicationsDetection Methods
      TFX, IF, ICCFluorescent
      Description
      Catalogue Number17-10228
      Trade Name
      • LentiBrite
      • Chemicon
      DescriptionLentiBrite™ Histone H2B-RFP Lentiviral Biosensor
      OverviewRead our application note in Nature Methods!
      http://www.nature.com/app_notes/nmeth/2012/121007/pdf/an8620.pdf
      (Click Here!)

      Learn more about the advantages of our LentiBrite Lentiviral Biosensors! Click Here

      Biosensors can be used to detect the presence/absence of a particular protein as well as the subcellular location of that protein within the live state of a cell. Fluorescent tags are often desired as a means to visualize the protein of interest within a cell by either fluorescent microscopy or time-lapse video capture. Visualizing live cells without disruption allows researchers to observe cellular conditions in real time.

      Lentiviral vector systems are a popular research tool used to introduce gene products into cells. Lentiviral transfection has advantages over non-viral methods such as chemical-based transfection including higher-efficiency transfection of dividing and non-dividing cells, long-term stable expression of the transgene, and low immunogenicity.

      EMD Millipore is introducing LentiBrite™ Lentiviral Biosensors, a new suite of pre-packaged lentiviral particles encoding important and foundational proteins of autophagy, apoptosis, and cell structure for visualization under different cell/disease states in live cell and in vitro analysis.
      • Pre-packaged, fluorescently-tagged with GFP & RFP
      • Higher efficiency transfection as compared to traditional chemical-based and other non-viral-based transfection methods
      • Ability to transfect dividing, non-dividing, and difficult-to-transfect cell types, such as primary cells or stem cells
      • Non-disruptive towards cellular function

      EMD Millipore’s LentiBrite™ Histone H2B-RFP lentiviral particles provide bright fluorescence and precise localization to enable live cell analysis of chromosomal dynamics in difficult-to-transfect cell types.
      Alternate Names
      • Histone H2B
      Background InformationChromatin, the higher order structure of DNA and nucleosomes, constitutes the majority of the nucleus of the eukaryotic cell. Changes in chromatin structure are the essence of many essential nuclear processes, including transcription, mitosis, meiosis, and apoptosis. The nucleosome comprises an octomer of four core histone proteins (H2A, H2B, H3 and H4). Genetic fusions between histone H2B and fluorescent proteins have been widely used in live cells to visualize the dynamics of chromosomal architecture during various processes. In addition to its utility for studying normal mitosis, histone H2B-GFP has been utilized to define mechanisms for asymmetric inheritance of oncogenic double minute chromosomes. Monitoring histone H2B-GFP also permits continuous analysis of chromosomal degradation during apoptosis. Use of histone H2B-GFP has recently been extended to live animals and high-throughput siRNA screens.
      EMD Millipore’s LentiBrite™ Histone H2B-RFP lentiviral particles provide bright fluorescence and precise localization to enable live cell analysis of chromosomal dynamics in difficult-to-transfect cell types.
      References
      Product Information
      Components
      • Histone H2B-TagRFP Lentivirus:
        One vial containing 25 µL of lentiviral particles at a minimum of 3 x 10E8 infectious units (IFU) per mL.
        For lot-specific titer information, please see lot specific “Viral Titer” in the product specifications of the datasheet.

      • Promoter
        EF-1 (Elongation Factor-1)

      • Multiplicty of Infection (MOI)
        MOI = Ratio of # of infectious lentiviral particles (IFU) to # of cells being infected.
        Typical MOI values for high transduction efficiency and signal intensity are in the range of 20-40. For this target, some cell types may require lower MOIs (e.g., HT-1080, HeLa, human mesenchymal stem cells (HuMSC)), while others may require higher MOIs (e.g., human umbilical vein endothelial cells (HUVEC), U2OS).
        NOTE: MOI should be titrated and optimized by the end user for each cell type and lentiviral target to achieve desired transduction efficiency and signal intensity.
      Detection methodFluorescent
      Quality LevelMQ100
      Applications
      Key Applications
      • Transfection
      • Immunofluorescence
      • Immunocytochemistry
      Application NotesFluorescence microscopy imaging:
      (See Figure 1 in datasheet)
      HeLa cells were plated in a chamber slide and transduced with lentiviral particles at an MOI of 20 for 24 hours. After media replacement and 48 hours further incubation, cells were fixed with formaldehyde and mounted. Images were obtained by oil immersion wide-field fluorescence microscopy. The histone H2B-RFP signal shows interphase nuclei, with the lower nuclei connected by a strand representing a segregation defect.

      Additional cell types and Hard-to-transfect cell types:
      (See Figure 2 in datasheet)
      Primary HUVECs were transduced at an MOI of 20. The image depicts an irregular interphase nucleus. HT1080 were transduced at an MOI of 40 for 24 hours. The image depicts an interphase nucleus. Human mesenchymal stem cells (HuMSC) transduced with lentiviral particles at an MOI of 40. The cell is in anaphase and displays lagging chromosomes.

      Mitosis in Various Stages:
      (See Figure 3 in datasheet)
      U2OS cells were plated in chamber slides and transduced with lentiviral particles at an MOI of 20 for 24 hours. The images show cells in various stages of mitosis.

      For optimal fluorescent visualization, it is recommended to analyze the target expression level within 24-48 hrs after transfection/infection for optimal live cell analysis, as fluorescent intensity may dim over time, especially in difficult-to-transfect cell lines. Infected cells may be frozen down after successful transfection/infection and thawed in culture to retain positive fluorescent expression beyond 24-48 hrs. Length and intensity of fluorescent expression varies between cell lines. Higher MOIs may be required for difficult-to-transfect cell lines.
      Biological Information
      Gene Symbol
      • H2B
      Purification MethodPEG precipitation
      UniProt Number
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Quality AssuranceEvaluated by transduction of HT-1080 cells and fluorescent imaging performed for assessment of target localization and transduction efficiency.
      Usage Statement
      • This product contains genetically modified organisms (GMO). Within the EU GMOs are regulated by Directives 2001/18/EC and 2009/41/EC of the European Parliament and of the Council and their national implementation in the member States respectively. This legislation obliges Merck to request certain information about you and the establishment where the GMOs are being handled. Click here for Enduser Declaration (EUD) Form.
      • 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.
      Storage and Shipping Information
      Storage ConditionsStorage and Handling
      Lentivirus is stable for at least 4 months from date of receipt when stored at -80°C. After first thaw, place immediately on ice and freeze in working aliquots at -80°C. Frozen aliquots may be stored for at least 2 months. Further freeze/thaws may result in decreased virus titer and transduction efficiency.

      IMPORTANT SAFETY NOTE
      Replication-defective lentiviral vectors, such as the 3rd Generation vector provided in this product, are not known to cause any diseases in humans or animals. However, lentiviruses can integrate into the host cell genome and thus pose some risk of insertional mutagenesis. Material is a Risk Group 2 and should be handled under BSL2 controls. A detailed discussion of biosafety of lentiviral vectors is provided in Pauwels, K. et al. (2009). State-of-the-art lentiviral vectors for research use: Risk assessment and biosafety recommendations. Curr. Gene Ther. 9: 459-474.
      Packaging Information
      Material Size1 vial (minimum of 3 x 10E8 IFU/mL)
      Transport Information
      Supplemental Information
      Specifications
      Global Trade Item Number
      Référence GTIN
      17-10228 04053252595936

      Documentation

      LentiBrite™ Histone H2B-RFP Lentiviral Biosensor FDS

      Titre

      Fiche de données de sécurité des matériaux (FDS) 

      LentiBrite™ Histone H2B-RFP Lentiviral Biosensor Certificats d'analyse

      TitreNuméro de lot
      LentiBrite Histone H2B-RFP Lentiviral Biosensor - Q1995263 Q1995263
      LentiBrite™ Histone H2B-RFP -2662280 2662280
      LentiBrite™ Histone H2B-RFP Lentiviral Biosensor 2438642
      LentiBrite™ Histone H2B-RFP Lentiviral Biosensor - 3466668 3466668
      LentiBrite™ Histone H2B-RFP Lentiviral Biosensor - 3520990 3520990
      LentiBrite™ Histone H2B-RFP Lentiviral Biosensor - 3726107 3726107
      LentiBrite™ Histone H2B-RFP Lentiviral Biosensor - 3886015 3886015
      LentiBrite™ Histone H2B-RFP Lentiviral Biosensor - 3932364 3932364
      LentiBrite™ Histone H2B-RFP Lentiviral Biosensor - 4076704 4076704
      LentiBrite™ Histone H2B-RFP Lentiviral Biosensor - 4113827 4113827
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      Brochure

      Titre
      New Products: Volume 3, 2012