Anti-DDDDK tag (Binds to FLAG® tag sequence) antibody [F-tag-01] (ab18230)

Overview

  • Product name

    Anti-DDDDK tag (Binds to FLAG® tag sequence) antibody [F-tag-01]
    See all DDDDK tag (Binds to FLAG® tag sequence) primary antibodies
  • Description

    Mouse monoclonal [F-tag-01] to DDDDK tag (Binds to FLAG® tag sequence)
  • Host species

    Mouse
  • Tested applications

    Suitable for: ICC, WB, ICC/IFmore details
    Unsuitable for: IP
  • Species reactivity

    Reacts with: Species independent
  • Immunogen

    Synthetic peptide corresponding to DDDDK tag conjugated to Keyhole Limpet Haemocyanin (KLH).
    Sequence:

    DYKDDDDK

  • General notes

    FLAG® is a registered trade mark of Sigma Aldrich Biotechnology LP.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.40
    Preservative: 0.097% Sodium azide
    Constituent: PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    F-tag-01
  • Isotype

    IgG1
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab18230 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC Use a concentration of 2 - 10 µg/ml.
WB Use a concentration of 2 µg/ml. Detects a band of approximately 55 kDa (predicted molecular weight: 55 kDa).
ICC/IF Use at an assay dependent dilution.
  • Application notes
    Is unsuitable for IP.
  • Target

    • Relevance

      This is a useful tool for the localisation and characterisation of DDDDK tagged proteins.
    • Alternative names

      • DDDDK epitope tag antibody
      • DDDK antibody
      • ddk antibody
      • DYKDDDDK antibody
      • DYKDDDDK epitope tag antibody
      • DYKDDDDK tag antibody
      • ECS epitope tag antibody
      • ECS tag antibody
      • Enterokinase Cleavage Site epitope tag antibody
      • Enterokinase Cleavage Site tag antibody
      • FLAG® antibody
      • FLAG® tag antibody
      see all

    Images

    • ab18230 staining DDDDK tag in Human HeLa, cervical carcinoma cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.3% tween20 and blocked with 0.5% BSA for 1 hour at 22°C. Samples were incubated with primary antibody (1/200 in PBS) for 24 hours at 4°C. Ab8517 (1/200) was used as the secondary antibody

      See Abreview

    • All lanes : Anti-DDDDK tag (Binds to FLAG® tag sequence) antibody [F-tag-01] (ab18230) at 2 µg/ml

      Lane 1 : Hek293 cells transfected with empty vector
      Lane 2 : HEK293 cells transfected with vector overexpressing DDDDK tag

      Secondary
      All lanes : Goat anti-mouse IgG conjugated to HRP

      Performed under reducing conditions.

      Predicted band size: 55 kDa



      Blocked for 16 hours/ 4oC in 3.0% Non-fat milk, PBS, 0.1% Tween-20

      See Abreview

    • (a) Fusion of nuclear protein stained with ab18230 (red)
      (b) Cell nuclei stained with DAPI (blue)
      (c) Merge of (a) and (b) confirming nuclear localisation of the fusion protein.
    • Immunofluorescence analysis of transfected HEK293 cells, staining DDDDK tag with ab18230 at 1/200 dilution.

    References

    This product has been referenced in:

    • Liang Q  et al. TRIM47 is up-regulated in colorectal cancer, promoting ubiquitination and degradation of SMAD4. J Exp Clin Cancer Res 38:159 (2019). Read more (PubMed: 30979374) »
    • Nehls J  et al. Release of Immunomodulatory Ebola Virus Glycoprotein-Containing Microvesicles Is Suppressed by Tetherin in a Species-Specific Manner. Cell Rep 26:1841-1853.e6 (2019). Read more (PubMed: 30759394) »
    See all 22 Publications for this product

    Customer reviews and Q&As

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    1-7 of 7 Abreviews

    Application
    Western blot
    Sample
    Human Purified protein (FLAG tagged purified SPOCK2 protein)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    0.001 µg
    Specification
    FLAG tagged purified SPOCK2 protein
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

    Abcam user community

    Verified customer

    Submitted Nov 19 2018

    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Monkey Cell (COS-7)
    Permeabilization
    Yes - 0.2% triton
    Specification
    COS-7
    Blocking step
    BSA as blocking agent for 40 minute(s) · Concentration: 5% · Temperature: 22°C
    Fixative
    Formaldehyde

    Abcam user community

    Verified customer

    Submitted Sep 12 2013

    Application
    Western blot
    Sample
    Mouse Recombinant protein (Rear leg muscle (homogenised))
    Gel Running Conditions
    Reduced Denaturing (16% Tricine)
    Loading amount
    20 µg
    Specification
    Rear leg muscle (homogenised)
    Blocking step
    Milk as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

    Abcam user community

    Verified customer

    Submitted Mar 04 2011

    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (293 cells transfected with empty vector or FH-FLAG)
    Gel Running Conditions
    Reduced Denaturing (Pierce precise protein gel 4-20%)
    Loading amount
    20 µg
    Specification
    293 cells transfected with empty vector or FH-FLAG
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

    Mr. Antibody Solutions

    Verified customer

    Submitted Dec 17 2010

    Application
    Western blot
    Sample
    Escherichia coli Cell lysate - whole cell (chimeric protein)
    Gel Running Conditions
    Reduced Denaturing (4-12% bis tris)
    Loading amount
    2 µg
    Specification
    chimeric protein
    Blocking step
    Milk as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

    Abcam user community

    Verified customer

    Submitted Nov 17 2010

    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HeLa, cervical carcinoma)
    Permeabilization
    Yes - 0,3% Tween20
    Specification
    HeLa, cervical carcinoma
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
    Fixative
    Paraformaldehyde

    Abcam user community

    Verified customer

    Submitted Mar 25 2009

    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HEK293 transfected with a 27 kDa construct)
    Loading amount
    25 µg
    Specification
    HEK293 transfected with a 27 kDa construct
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3%

    Abcam user community

    Verified customer

    Submitted Apr 18 2007

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