Anti-DDDDK tag antibody - (Equivalent to FLAG antibodies from Sigma) (ab1162)

Overview

  • Product nameAnti-DDDDK tag antibody - (Equivalent to FLAG antibodies from Sigma)
    See all DDDDK tag primary antibodies
  • Description
    Rabbit polyclonal to DDDDK tag - (Equivalent to FLAG antibodies from Sigma)
  • Tested applicationsWB, ICC/IF, ICC, ELISAmore details
  • Immunogen

    Peptide

    xxxDDDDK

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

  • General notesEpitope 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

  • FormLiquid
  • Storage instructionsStore at +4°C short term (1-2 weeks). Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage bufferPBS with 0.1% sodium azide
  • Concentration information loading...
  • PurityImmunogen affinity purified
  • Clonality Polyclonal
  • IsotypeIgG
  • 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.
ICC Use at an assay dependent concentration.
ELISA Use a concentration of 1 - 5 µg/ml.

as coating antibody in sELISA.

Target

  • RelevanceThis 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

Anti-DDDDK tag antibody - (Equivalent to FLAG antibodies from Sigma) 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 for Anti-DDDDK tag antibody - (Equivalent to FLAG antibodies from Sigma) (ab1162)

This product has been referenced in:

See all 20 Publications for this product

Product Wall

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...

Read More

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 ...

Read More

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...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (HeLA)
Loading amount 25 µg
Specification HeLA
Gel Running Conditions Reduced Denaturing (10)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

Submitted Jan 21 2012

Thank you very much for getting back to me so quickly. There seems to be some discrepancy which may cause the problem. The primary antibody (ab1162) is rabbit polyclonal to DDDDK tag and the secondary antibody is goat anti-mouse. The primary and th...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Recombinant protein (Flag-tagged recombinant protein)
Loading amount 2 µg
Specification Flag-tagged recombinant protein
Gel Running Conditions Reduced Denaturing (4-20% Tris-Glycine Gel)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Abcam user community

Verified customer

Submitted Aug 06 2010

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