Recombinant
RabMAb

Recombinant Anti-DDDDK tag antibody [EPR20018-251] - (Equivalent to FLAG antibodies from Sigma) (ab205606)

Overview

  • Product name

    Anti-DDDDK tag antibody [EPR20018-251] - (Equivalent to FLAG antibodies from Sigma)
    See all DDDDK tag primary antibodies
  • Description

    Rabbit monoclonal [EPR20018-251] to DDDDK tag - (Equivalent to FLAG antibodies from Sigma)
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ICC/IF, Flow Cyt, IHC-P, IPmore details
  • Species reactivity

    Reacts with: Species independent
  • Immunogen

    Synthetic peptide within Human DDDDK tag. The exact sequence is proprietary.

  • Positive control

    • WB: HEK-293 transfected with PFKFB3 and HEK-293 transfected with PD-L1 cell lysate. IHC-P: HEK-293T transfected with DDDDK-tagged human PD-L1. ICC/IF: HEK-293T cells. Flow: HEK-293T transfected with DDDDK-tagged human PD-L1. IP: HEK-293T transfected with DDDDK-tagged human PFKFB3.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

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 long term. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.01% Sodium azide
    Constituents: PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR20018-251
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab205606 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
WB 1/5000.
ICC/IF 1/100.
Flow Cyt 1/700.
IHC-P 1/750.
IP 1/30.

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

  • Immunhistochemical analysis of agarose-embedded HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with DDDDK-tagged human PD-L1 expression vector labeling DDDDK tag with ab205606 at 1/750 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Counterstained with hematoxylin.

    Positive staining on HEK-293T cells transfected with DDDDK-tagged human PD-L1 expression vector (Panel A) is observed. No signal was detected on HEK-293T transfected with an empty vector (vector control), containing a C-terminal DDDDK tag (Panel B).

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) cells labeling DDDDK tag with ab205606 at 1/100 dilution, followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution (green).

    Confocal image showing positive staining for Flag on HEK-293T cells transfected with DDDDK-tagged PFKFB3 expression vector. Mouse monoclonal ANTI-FLAG® M2 antibody was used as a counterstain at 1/500 dilution, and AlexaFluor®647 Goat anti-mouse secondary (ab150115) was used as the secondary antibody only control at 1/500 dilution. The nucleus is counterstained with DAPI.

  • All lanes : Anti-DDDDK tag antibody [EPR20018-251] - (Equivalent to FLAG antibodies from Sigma) (ab205606) at 1/1000 dilution

    Lane 1 : HEK-293 (Human epithelial cell line from embryonic kidney) transfected with an empty vector (vector control), containing an N-terminal DDDDK tag, whole cell lysate
    Lane 2 : HEK-293 transfected with PFKFB3 (WT) expression vector containing an N-terminal DDDDK tag, whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Observed band size: 60 kDa
    why is the actual band size different from the predicted?



    Blocking/Diluting buffer: 5% NFDM/TBST 

    Exposure time: 0.5 seconds

  • DDDDK tag was immunoprecipitated from 0.35 mg HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with DDDDK-tagged human PFKFB3 expression vector whole cell lysate with ab205606 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab205606 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: HEK-293T transfected with DDDDK-tagged human PFKFB3 expression vector whole cell lysate (input).

    Lane 2: ab205606 IP in HEK-293T transfected with DDDDK-tagged human PFKFB3 expression vector whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab205606 in HEK-293T transfected with DDDDK-tagged human PFKFB3 expression vector whole cell lysate.

    Blocking/Dilution buffer: 5% NFDM/TBST
    Exposure time: 3 seconds. 

  • Flow cytometric analysis of of 4% paraformaldehyde-fixed, 90% methanol permeabilized HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with DDDDK-tagged human PD-L1 expression vector labeling DDDDK tag with ab205606 at 1/700 dilution (Red) compared with the Rabbit monoclonal IgG isotype control (ab172730) (Black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077), at 1/2000 dilution was used as the secondary antibody.

  • Negative control: No staining on rat spleen.

    Immunohistochemical analysis of paraffin-embedded rat spleen tissue stained for DDDDK tag using ab205606 at 1/750 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Counterstained with hematoxylin.

  • Negative control: No staining on mouse spleen.

    Immunohistochemical analysis of paraffin-embedded mouse spleen tissue stained for DDDDK tag using ab205606 at 1/750 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Counterstained with hematoxylin.

  • Negative control: No staining on human hepatocellular cancer.

    Immunohistochemical analysis of paraffin-embedded human hepatocellular cancer tissue stained for DDDDK tag using ab205606 at 1/750 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Counterstained with hematoxylin.

  • Lane 1 : Anti-DDDDK tag antibody [EPR20018-251] - (Equivalent to FLAG antibodies from Sigma) (ab205606) at 1/5000 dilution
    Lane 2 : Anti-DDDDK tag antibody [EPR20018-251] - (Equivalent to FLAG antibodies from Sigma) (ab205606) at 1/100000 dilution

    Lane 1 : HEK-293 (Human epithelial cell line from embryonic kidney) transfected with an empty vector (vector control), containing a C-terminal DDDDK tag, whole cell lysate
    Lane 2 : HEK-293 transfected with PD-L1 (WT) expression vector containing a C- terminal DDDDK tag, whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Lane 1 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000000 dilution
    Lane 2 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution


    Blocking/Diluting buffer: 5% NFDM/TBST

    Exposure time: 1 second

References

This product has been referenced in:

  • Li Z  et al. GDNF family receptor alpha 2 promotes neuroblastoma cell proliferation by interacting with PTEN. Biochem Biophys Res Commun 510:339-344 (2019). Read more (PubMed: 30722993) »
  • Xu W  et al. Endothelial FAM3A positively regulates post-ischaemic angiogenesis. EBioMedicine 43:32-42 (2019). Read more (PubMed: 31000420) »
See all 3 Publications for this product

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