Flavivirus E protein antibody

Details for Product No. ABIN3200993, Supplier: Log in to see
Antigen
Reactivity
Human, Mouse (Murine)
Host
Mouse
Clonality (Clone)
Monoclonal ()
Application
Immunofluorescence (IF), Western Blotting (WB)
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'Independent Validation' Badge
Antigen Flavivirus E protein
Method validated Immunocytochemistry
Positive Control 1205Lu human melanoma cells infected with yellow fever vaccine strain YF-17D
Negative Control Uninfected 1205Lu cells; infected cells without primary antibody
Notes 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.
Primary Antibody ABIN3200993
Secondary Antibody goat-anti mouse AF647 conjugated antibody (Invitrogen, A21235, lot 1764240)
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.
Validation Images
Immunocytochemistry validation image for anti-Flavivirus E protein antibody (ABIN3200993) 1205Lu melanoma cells infected (B, C, D) and uninfected (A) with yellow fever vaccine...
Immunogen Dengue Virus
Clone 4G2
Isotype IgG2a
Specificity E protein of Flavivirus
Characteristics 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).  
Purification Purified from hybridoma supernatant via Protein G
Application Notes 0.1-1.0 µg/mL
Restrictions For Research Use only
Format Liquid
Concentration 1 mg/mL
Buffer PBS pH 7.4
Storage -20 °C
Storage Comment Heat stable , shipped at ambient  temp  Upon delivery aliquot and store in fridge , longterm storage at  -20°C.
Supplier Images
Immunofluorescence (IF) image for anti-Flavivirus E protein antibody (ABIN3200993) IF of Zika infected cells
Product cited in: 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).

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