Overview

  • Product name
  • Description
    Rabbit polyclonal to DDDDK tag
  • Tested applications
    Suitable for: WB, ICC/IF, ELISA, IPmore details
  • Immunogen

    Peptide xxxDDDDK conjugated to KLH. xxx represents random amino acids. This sequence represents the Enterokinase Cleavage Site.

  • General notes
    Epitope tags provide a method to localize gene products in a variety of cell types, study the topology of proteins and protein complexes, identify associated proteins, and characterize newly identified, low abundance or poorly immunogenic proteins when protein specific antibodies are not available. Tagging with xxxDDDDK may be done at the N-terminus, N-terminus preceded by a methionine residue, C-terminus, and in internal positions of the target protein. The small size of the epitope tag and its high hydrophilicity tend to decrease the possibility of interference with protein expression, proteolytic maturation, antigenicity and function. The enterokinase cleavage site allows it to be completely removed from the purified fusion proteins.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer
    PBS with 0.1% sodium azide
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab1162 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 Use a concentration of 0.2 - 1 µg/ml.

Antibody was analyzed by western blot using various amino-terminal and carboxy-terminal DDDDK fusion proteins and Invitrogen Positope" R900-40. A 1/5000 dilution gave bands against 0.1, 1.0 and 10 ng of the fusion proteins and 100 ng and 25 ng of the Positope.

ICC/IF Use at an assay dependent concentration.
ELISA Use a concentration of 1 - 5 µg/ml.

As coating antibody in sELISA.

IP 1/40. PubMed: 25419851

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

  • ab1162 staining DDDDK tag in hTERT-RPE1 by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with Paraformaldehyde, permeabilized with Saponin and blocked with 0.02% BSA for 20 minutes at 18°C. Samples were incubated with primary antibody (1/100) for 45 minutes at 18°C. A Cy3®-conjugated Donkey anti-rabbit polyclonal was used as the secondary antibody.

    See Abreview

  • ab1162 staining DDDDK tag (pink) in neonatal rat cardiomyocytes transduced with Ad-TBX18/FLAG-IRES-GFP by Immunocytochemistry/ Immunofluorescence.

    Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton X-100 and incubated with primary antibody. An AlexaFluor®-conjugated anti-rabbit IgG was used as the secondary antibody. Nuclei were stained with DAPI (blue).

References

This product has been referenced in:

See all 48 Publications for this product

Customer reviews and Q&As

Application
Western blot
Sample
Mouse Cell lysate - other (neuroblastoma2 a cells)
Gel Running Conditions
Reduced Denaturing (10)
Loading amount
10 µg
Specification
neuroblastoma2 a cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Mar 29 2017

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (brain)
Permeabilization
Yes - Triton x-100
Specification
brain
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2.5% · Temperature: 25°C
Fixative
Paraformaldehyde
Username

久仁子 島崎

Verified customer

Submitted Sep 05 2016

Application
Western blot
Sample
Human Cell lysate - whole cell (HeLa)
Gel Running Conditions
Reduced Denaturing (10%)
Loading amount
10 µg
Specification
HeLa
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Apr 22 2016

Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 20°C
Antigen retrieval step
None
Sample
Mouse Tissue sections (Brain- striatal medium spiny neurons)
Specification
Brain- striatal medium spiny neurons
Permeabilization
Yes - 0.03% Triton
Fixative
10% Formalin
Username

Rivka Schwarcz

Verified customer

Submitted Jul 28 2014

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Sample
Monkey Cell (COS7)
Specification
COS7
Permeabilization
Yes - Tritonx 100
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

Submitted Oct 18 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (hTERT-RPE1)
Specification
hTERT-RPE1
Fixative
Paraformaldehyde
Permeabilization
Yes - saponin
Blocking step
BSA as blocking agent for 20 minute(s) · Concentration: 0.02% · Temperature: 18°C
Username

Abcam user community

Verified customer

Submitted Dec 19 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (hTERT-RPE1)
Loading amount
50000 cells
Specification
hTERT-RPE1
Treatment
transfected with flag tag plasmid 20hr
Gel Running Conditions
Reduced Denaturing (8 % acrylamid)
Blocking step
Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Abcam user community

Verified customer

Submitted Dec 14 2012

Merci de nous avoir contactés.

Nous avons plusieurs anti-FLAG anticorps au catalogue par contre aucun n’a été testé en microscopie électronique.

Cependant, je peux vous proposer :

a...

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Gracias por tu confirmacion.


Acabo de hacer un nuevo pedido del anticuerpo ab1259 de forma totalmente gratuita. El número de pedido es XXXX. A lo largo del día recibirás una confirmación automática del ...

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Te agradezco mucho que nos hayas facilitado mas detalles del protocolo y te hayas tomado la molestia de enviar las imágenes.

Es evidente que el anticuerpo no es especifico contra el DDDDK tag visto que da la misma señal en c&eac...

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1-10 of 21 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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