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DYKDDDDK Epitope Tag Antibody [Janelia Fluor® 525]

 
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Product Details

Summary
Reactivity ETSpecies Glossary
Applications WB, Simple Western, ELISA, ICC/IF, IHC, IP
Clonality
Polyclonal
Host
Rabbit
Conjugate
Janelia Fluor 525

Order Details

View Available Conjugates
Catalog# & Conjugate Size Price
Alexa Fluor 350
NB600-345AF350
Alexa Fluor 405
NB600-345AF405
Alexa Fluor 488
NB600-345AF488
Alexa Fluor 532
NB600-345AF532
Alexa Fluor 594
NB600-345AF594
Alexa Fluor 647
NB600-345AF647
Alexa Fluor 700
NB600-345AF700
Alexa Fluor 750
NB600-345AF750
Allophycocyanin
NB600-345APC
Biotin
NB600-345B
CoraFluor 1
NB600-345CL1
DyLight 350
NB600-345UV
DyLight 405
NB600-345V
DyLight 488
NB600-345G
DyLight 550
NB600-345R
DyLight 594
NB600-345DL594
DyLight 650
NB600-345C
DyLight 680
NB600-345FR
DyLight 755
NB600-345IR
FITC
NB600-345F
HRP
NB600-345H
Janelia Fluor 549
NB600-345JF549
Janelia Fluor 585
NB600-345JF585
Janelia Fluor 635
NB600-345JF635
Janelia Fluor 646
NB600-345JF646
Janelia Fluor 669
NB600-345JF669
PE
NB600-345PE
PerCP
NB600-345PCP
Unconjugated
NB600-345
View Available Formulations
Catalog# & Formulation Size Price
Standard Formulation
NB600-345
BSA Free
NB600-345
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DYKDDDDK Epitope Tag Antibody [Janelia Fluor® 525] Summary

Immunogen
To produce DYKDDDDK Epitope Tag Antibody, Rabbits were immunized with ECS enterokinase cleavage site (xxxDDDDK) conjugated to KLH. Antibody was isolated by affinity chromatography using the peptide immobilized on solid support.
Specificity
DYKDDDDK Epitope Tag Antibody reacts with FLAG.
Isotype
IgG
Clonality
Polyclonal
Host
Rabbit
Purity
Immunogen affinity purified
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Applications/Dilutions

Dilutions
  • ELISA
  • Immunocytochemistry/ Immunofluorescence
  • Immunohistochemistry
  • Immunoprecipitation
  • Simple Western
  • Western Blot
Application Notes
Optimal dilution of this antibody should be experimentally determined.

Packaging, Storage & Formulations

Storage
Store at 4C in the dark.
Buffer
50mM Sodium Borate
Preservative
0.05% Sodium Azide
Purity
Immunogen affinity purified

Notes



Sold under license from the Howard Hughes Medical Institute, Janelia Research Campus.

Alternate Names for DYKDDDDK Epitope Tag Antibody [Janelia Fluor® 525]

  • DDDDK epitope tag
  • DDK
  • DYKDDDDK Epitope Tag
  • DYKDDDDK tag
  • ECS epitope tag
  • ECS tag
  • Enterokinase Cleavage Site epitope tag
  • Enterokinase Cleavage Site tag
  • epitope tag flag
  • FLAG (FLAG is a trademark of Sigma Aldrich)
  • FLAG Tag
  • Ty 1 Tag
  • xxxDDDDK epitope tag

Background

Fusion proteins, in which a known peptide such as a His-Tag, GST-Tag, or DYKDDDDK (FLAG(R)) is cloned with the target of interest, were developed to simplify protein isolation. Since specific antibodies may not be available against the protein of interest, epitope tag antibodies are commonly used to detect protein expression.

The epitope tag, DYKDDDDK, is comprised of an eight amino acid sequence derived from the gene-10 product from bacteriophage T7 and contains an enterokinase-cleavage site (1). Its hydrophilic nature and short peptide sequence reduce possible interference with protein expression, post-translational modifications, protein interactions, and protein function (2). The DYKDDDDK (FLAG(R)) epitope tag can either be fused to the N-terminus, C-terminus, or placed at an internal position of the target protein, facilitating characterization of the protein of interest using immunodetection methods. Compared to the standard DYKDDDDK epitope tag antibody, the rat monoclonal DYKDDDDK Epitope Tag antibody (clone L5) is 2-8 times more sensitive (3).

References

1. Brizzard BL, Chubet RG, Vizard DL. (1994) Immunoaffinity purification of (FLAG(R)) epitope-tagged bacterial alkaline phosphatase using a novel monoclonal antibody and peptide elution. Biotechniques. 16(4):730-5. PMID: 8024796

2. Einhauer A, Jungbauer A. (2001) The (FLAG(R)) peptide, a versatile fusion tag for the purification of recombinant proteins. J Biochem Biophys Methods. 49(1-3):455-65. PMID: 11694294

3. Park SH, Cheong C, Idoyaga J, Kim JY, Choi JH, Do Y, Lee H, Jo JH, Oh YS, Im W, Steinman RM, Park CG. (2008) Generation and application of new rat monoclonal antibodies against synthetic DYKDDDDK Epitope Tag (FLAG(R)) and OLLAS tags for improved immunodetection. J Immunol Methods. 331(1-2):27-38. PMID: 18054954

FLAG(R) is a trademark of Sigma-Aldrich.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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Product General Protocols

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

Video Protocols

WB Video Protocol
ICC/IF Video Protocol

FAQs for DYKDDDDK Epitope Tag Antibody (NB600-345JF525). (Showing 1 - 5 of 5 FAQ).

  1. Are the FLAG Tag antibodies validated in Simple Western?
    • Yes, we offer 3 FLAG Tag antibodies that have been tested in Simple Western: NBP1-71705, NBP1-97410, and MAB8529.
  2. What the theoretical molecular weight for FLAG Tag antibodies?
    • The TMW for DYKDDDDK epitope FLAG Tag is approximately 1.01 kDa.
  3. Will FLAG Tag antibodies detect internal FLAG tags? My FLAG sequence is sandwiched between two other protein sequences.
    • If you are applying your sample(s) to SDS-PAGE, under reducing condition and follow by Western Blot, DYKDDDDK epitope tag antibodies should be able to detect the tag region located internally. However, it depends on the property of your chimeric protein and if your tag is in the hydrophobic or hydrophilic region. Although DYKDDDDK is very hydrophilic, it may not be exposed to the surface if embedded deeply inside the protein with a high content of hydrophobic amino acids with or without covalently-linked modifications such as lipid moieties.
  4. Will FLAG Tag antibodies recognize FLAG at the C-terminal end?
    • Yes, FLAG Tag antibodies can recognize FLAG-tag at both the N- or C-terminal.
  5. Do FLAG Tag antibodies comes in lyophilized form?
    • The following FLAG Tag antibodies come in lyophilized form: NBP1-97393, NBP1-97401, NBP1-97399, NBP1-97400, and NBP1-97402.

Secondary Antibodies

 

Isotype Controls

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