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DYKDDDDK Tag antibody

WB, ELISA, IP, DB, ISt Host: Mouse Monoclonal FG4R unconjugated
Catalog No. ABIN559688
  • Target See all DYKDDDDK Tag products
    DYKDDDDK Tag
    Reactivity
    Please inquire
    Host
    • 57
    • 52
    • 8
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    Mouse
    Clonality
    • 65
    • 63
    Monoclonal
    Conjugate
    • 45
    • 15
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    • 3
    • 2
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    This DYKDDDDK Tag antibody is un-conjugated
    Application
    • 79
    • 61
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    Western Blotting (WB), ELISA, Immunoprecipitation (IP), Dot Blot (DB), Immunostaining (ISt)
    Sequence
    DYKDDDDK
    Specificity
    Recognizes the N-terminal, C-terminal or internal DYKDDDDK-tagged fusion proteins
    Sensitivity
    WB (with ECL): 1-5ng of DYKDDDDK-tagged fusion proteins
    Purification
    Protein A affinity chromatography from mouse ascites fluid
    Immunogen
    DYKDDDDK (DYKDDDDK) synthetic peptide conjugated to KLH
    Clone
    FG4R
    Isotype
    IgG1
  • Application Notes
    WB (with ECL): 1:1000-3000 dilution (incubate for one hour at room temperature) / IS: 1:500-2000 dilution / For best results with other assays (e.g.: Dot, ELISA, IP, etc), please determine optimal working dilution by titration test
    Restrictions
    For Research Use only
  • Validation #102796 (Immunofluorescence)
    'Independent Validation' Badge
    by
    Martinelli lab, Neuroscience Department, UConn Health
    No.
    #102796
    Date
    02/13/2018
    Antigen
    DYKDDDDK-tag
    Lot Number
    02017
    Method validated
    Immunofluorescence
    Positive Control
    African green monkey kidney fibroblast-like COS7 cells transfected with BAI3, a transmembrane protein with a surface presented DYKDDDDK-tag
    Negative Control
    untransfected COS7 cells
    Notes

    Passed. Signal was observed as expected. There was never any signal on untransfected cells, demonstrating specificity of the antibody.

    'Independent Validation' Badge
    Validation Images
    Full Methods
    Primary Antibody
    ABIN1715174
    Secondary Antibody
    goat anti-mouse AF488 conjugated antibody (Life Technologies, A11029, lot 1829920)
    Full Protocol
    • Grow COS7 cells in DMEM (Biological Industries, 01-052-1A, lot 1728619) supplemented with FBS (Hyclone, SH3039602) without antibiotics, at 37°C and 5% CO2 in 1ml on glass coverslips on 24-well plate.
    • Transfect cells with calcium phosphate method with a eukaryotic expression construct to express a DYKDDDDK-tagged BAI3, which will be surface presented.
    • Grow cells for 24h.
    • Fix cells on coverslips in 4% PFA for 15min at 4°C.
    • Wash cells 3x for 5min with PBS.
    • Block non-specific binding with 5% goat serum in PBS for 1h at RT.
    • Incubate cells with primary mouse anti-DYKDDDDK-tag antibody (antibodies-online, ABIN1715174, lot 02017) diluted 1:2000 in blocking solution ON at 4°C.
    • Wash cells 3x for 5min with PBS.
    • Incubate cells with secondary goat anti-mouse AF488 conjugated antibody (Life Technologies, A11029, lot 1829920) diluted 1:400 in blocking buffer for 1h at RT protected from light.
    • Wash cells 3x for 5min with PBS.
    • Mount coverslips on glass slides in Fluoromount-G with DAPI mounting medium (ThermoFisher Scientific, 00-4959-52, lot B2215-N915).
    • Image acquisition on a Zeiss Axiovert epifluorescence with with a 40x objective, using a Zeiss ApoTome for optical sectioning.
    Experimental Notes
  • Concentration
    1mg/mL
    Buffer
    10mM PBS, pH 7.2, 0.05% NaN3
    Preservative
    Sodium azide
    Precaution of Use
    This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
    Storage
    -20 °C
  • Target
    DYKDDDDK Tag
    Alternative Name
    FLAG Tag (DYKDDDDK Tag Products)
    Target Type
    Tag
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