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Flavivirus E protein Antikörper

Reaktivität: Human, Maus IF, WB Wirt: Maus Monoclonal 4G2 unconjugated
Produktnummer ABIN3200993
  • Target
    Flavivirus E protein
    Reaktivität
    Human, Maus
    Wirt
    Maus
    Klonalität
    Monoklonal
    Applikation
    Immunofluorescence (IF), Western Blotting (WB)
    Spezifität
    E protein of Flavivirus
    Produktmerkmale
    Mouse monclonal generated against flavivirus envelope proteins.  Recognizes Dengue virus, West Nile Virus, Japanese Encephalitis and  Zika virus. It binds to domain II of protein E (fusion loop).  
    Aufreinigung
    Purified from hybridoma supernatant via Protein G
    Immunogen
    Dengue Virus
    Klon
    4G2
    Isotyp
    IgG2a
  • Applikationshinweise
    0.1-1.0 µg/mL
    Beschränkungen
    Nur für Forschungszwecke einsetzbar
  • Validierung #101139 (Immunocytochemistry)
    'Independent Validation' Siegel
    by
    Division of Clinical Pharmacology, LMU, Munich
    No.
    #101139
    Datum
    07.11.2017
    Antigen
    Flavivirus E protein
    Chargennummer
    Validierte Anwendung
    Immunocytochemistry
    Positivkontrolle
    1205Lu human melanoma cells infected with yellow fever vaccine strain YF-17D
    Negativkontrolle
    Uninfected 1205Lu cells; infected cells without primary antibody
    Bewertung
    ABIN3200993 specifically labels the targeted antigen in 1205Lu melanoma cells in ICC. No signal was detected in uninfected negative control tissue and the secondary antibody only control.
    'Independent Validation' Siegel
    Validierungsbilder
    Protokoll
    Primärantikörper
    ABIN3200993
    Sekundärantikörper
    goat-anti mouse AF647 conjugated antibody (Invitrogen, A21235, lot 1764240)
    Full Protocol
    • Grow cells in DMEM supplemented with 10% FCS, 1% L-Glutamin, 1% Pen/Strep (Gibco) to 70% confluency on coverslips coated with Poly L-Lysin (Sigma, P8920).
    • Infect cells with yellow fever vaccine strain YF-17D at 2MOI.
    • Grow cells on coverslips for 48h.
    • Fix cells on coverslips in 4% PFA for 30min at RT.
    • Wash cells 2x with PBS.
    • Permeabilize cells in PBScontaining 0.1% Triton-X 100 (PBST) for 1h at RT.
    • Wash cells 2x with PBS.
    • Block non-specific binding with in PBST containing 10% normal goat serum for 2h at RT.
    • Incubate slides with primary mouse anti-Flavivirus E protein antibody (antibodies-online, ABIN3200993) diluted 1:200 and rabbit-anti ER antibody (abcam, ab176333, GR265198-3) diluted 1:200 in PBST with 10% NGS ON at 4°C.
    • Include wells without primary antibody as negative controls.
    • Rinse slides 3x with PBST.
    • Incubate slides with secondary goat-anti mouse AF647 conjugatedantibody (Invitrogen, A21235, lot 1764240) diluted 1:1000 and goat-anti rabbit AF488 conjugated antibody (Invitrogen, A-11008, lot 84B2-1) diluted 1:1000 in PBST with 3% NGS for 1h at RT.
    • Rinse slide 3x with PBST.
    • Nuclear counterstain with DAPI diluted in PBS (Sigma, 10236276001).
    • Wash cells 2x with PBS.
    • Mount coverslips on glass slides in Mowiol antifade reagent (Calbiochem, 475904).
    • Image acquisition with Leica SP5 II confocal imaging system, 20x magnification, 1024x1024 resolution.
    Anmerkungen
  • Format
    Liquid
    Konzentration
    1 mg/mL
    Buffer
    PBS pH 7.4
    Lagerung
    -20 °C
    Informationen zur Lagerung
    Heat stable , shipped at ambient  temp  Upon delivery aliquot and store in fridge , longterm storage at  -20°C.
  • Tang, Hammack, Ogden, Wen, Qian, Li, Yao, Shin, Zhang, Lee, Christian, Didier, Jin, Song, Ming: "Zika Virus Infects Human Cortical Neural Progenitors and Attenuates Their Growth." in: Cell stem cell, Vol. 18, Issue 5, pp. 587-90, (2017) (PubMed).

  • Target
    Flavivirus E protein
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