DYKDDDDK Epitope Tag Antibody

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Western Blot: DYKDDDDK Epitope Tag Antibody [NB600-344] - Samples: 200, 100, or 50 ng of E. coli whole cell lysate expressing a multi-tag fusion protein. Antibody: Affinity purified goat DYKDDDDK Epitope Tag Antibody ...read more
Western Blot: DYKDDDDK Epitope Tag Antibody [NB600-344] - Analysis using the Agarose Immobilized conjugate of DYKDDDDK Epitope Tag Antibody [NB600-344]. DYKDDDK in transfected COS-7 cells using agarose immobilized ...read more
Immunocytochemistry/ Immunofluorescence: DYKDDDDK Epitope Tag Antibody [NB600-344] - Expression of the ZFN as detected by PLA.PK-15 cells were transfected with the pZFN1 or pZFN2 vectors expressing the 3xFLAG tagged ...read more
Immunocytochemistry/ Immunofluorescence: DYKDDDDK Epitope Tag Antibody [NB600-344] - Samples: Human microvascular endothelial cells expressing FLAG tagged beta-catenin following transient transfection. Antibody: ...read more

Product Details

Summary
Reactivity Hu, All-NA, Pm, PmSpecies Glossary
Applications WB, ELISA, ICC/IF, IP
Clonality
Polyclonal
Host
Goat
Conjugate
Unconjugated
Concentration
1.0 mg/ml

DYKDDDDK Epitope Tag Antibody Summary

Immunogen
To produce DYKDDDDK Epitope Tag Antibody, Goats 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
Goat
Purity
Immunogen affinity purified
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.

Applications/Dilutions

Dilutions
  • Western Blot 1:1000-1:30000
  • ELISA 1:100-1:500 (coating); 1:1000-1:30000 (primary)
  • Immunocytochemistry/Immunofluorescence 1:100-1:400
  • Immunoprecipitation 1-4 ug/mg Lysate
Application Notes
Use in IP, WB, and ICC/IF reported in multiple pieces of scientific literature.
Theoretical MW
1.01 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Publications
Read Publications using
NB600-344 in the following applications:

  • 2 publications
  • IP
    2 publications
  • PLA
    1 publication
  • WB
    6 publications

Reactivity Notes

Human and Primate reactivity reported in multiple pieces of scientific literature. Monkey reactivity reported in scientific literature (PMID: 26493046).

Packaging, Storage & Formulations

Storage
Store at 4C. Do not freeze.
Buffer
PBS
Preservative
0.09% Sodium Azide
Concentration
1.0 mg/ml
Purity
Immunogen affinity purified

Notes

FLAG(R) and ANTI-FLAG(R) are registered trademarks of Sigma-Aldrich Co. LLC.

Alternate Names for DYKDDDDK Epitope Tag Antibody

  • 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.

Publications for DYKDDDDK Epitope Tag Antibody (NB600-344)(19)

We have publications tested in 4 confirmed species: Human, Monkey, Porcine, Primate.

We have publications tested in 4 applications: ICC/IF, IP, PLA, WB.


Filter By Application
ICC/IF
(2)
IP
(2)
PLA
(1)
WB
(6)
All Applications
Filter By Species
Human
(4)
Monkey
(1)
Porcine
(1)
Primate
(2)
All Species
Showing Publications 1 - 10 of 19. Show All 19 Publications.
Publications using NB600-344 Applications Species
Morii M, Kubota S, Honda T et al. Src Acts as an Effector for Ku70-dependent Suppression of Apoptosis through Phosphorylation of Ku70 at Tyr-530. J. Biol. Chem. Dec 20 2016 [PMID: 27998981] (WB) WB
Yun HJ, Park J, Ho DH et al. LRRK2 phosphorylates Snapin and inhibits interaction of Snapin with SNAP-25 Exp Mol Med Jul 5 1905

Details:
This publication used the Agarose Immobilized conjugated form of this antibody (Cat# NB600-351).
Shutt T, Zhao T, Goedhart C et al. PISD is a mitochondrial disease gene causing skeletal dysplasia, cataracts, and white matter changes Life Sci Alliance. [PMID: 30858161] (WB, Human) WB Human
Yang Y, Qin M, Bao P et al. Secretory carrier membrane protein 5 is an autophagy inhibitor that promotes the secretion of alpha-synuclein via exosome PLoS ONE 2017 Jul 12 [PMID: 28700687] (WB, Human) WB Human
Gorur A, Yuan L, Kenny SJ et al. COPII-coated membranes function as transport carriers of intracellular procollagen-1. bioRxiv. Feb 21 2017 [PMID: 28428367] (ICC/IF, WB, Human) ICC/IF, WB Human
Yokoi S, Ishihara N, Miya F et al. TUBA1A mutation can cause a hydranencephaly-like severe form of cortical dysgenesis. Sci Rep. 2015 Oct 23 [PMID: 26493046] (WB, Monkey) WB Monkey
Semaan M, Ivanusic D, Denner J. Cytotoxic Effects during Knock Out of Multiple Porcine Endogenous Retrovirus (PERV) Sequences in the Pig Genome by Zinc Finger Nucleases (ZFN) PLoS One 2015 Apr 24 [PMID: 25909512] (PLA, Porcine) PLA Porcine
Mallilankaraman K, Cardenas C, Doonan PJ et al. MCUR1 is an essential component of mitochondrial Ca(2+) uptake that regulates cellular metabolism Nat Cell Biol (IP, Primate)

Details:
This publication used the Agarose Immobilized conjugated form of this antibody (Cat# NB600-351).
IP Primate
Yun HJ, Park J, Ho DH et al. LRRK2 phosphorylates Snapin and inhibits interaction of Snapin with SNAP-25 Exp Mol Med 2013 [PMID: 23949442]

Details:
This citation used the Agarose Immobilized version of this antibody.
Mallilankaraman K, Cardenas C, Doonan PJ et al. MCUR1 is an essential component of mitochondrial Ca(2+) uptake that regulates cellular metabolism Nat Cell Biol 2012 Dec [PMID: 23178883] (IP, Primate)

Details:
This citation used the Agarose Immobilized version of this antibody.
IP Primate
Show All 19 Publications.

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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 (NB600-344). (Showing 1 - 7 of 7 FAQs).

  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.
  6. Is this antibody suitable for ChIP analysis of an N-terminally tagged protein?
    • Our DYKDDDDK epitope tag antibody with catalogue number NB600-344 has been validated for Western blotting, immunofluorescence, immunoprecipitation and ELISA but has not yet been tested for ChIP. However, our DYKDDDDK Epitope Tag antibody with catalog number NBP2-59146 is validated for use in ChIP.
  7. What is the immunogen sequence of this FLAG Tag antibody? 
    • This antibody was made to a ECS enterokinase cleavage site (xxxDDDDK) conjugated to KLH.  

Secondary Antibodies

 

Isotype Controls

Other Available Formats

Agarose NB600-351
Alexa Fluor 350 NB600-344AF350
Alexa Fluor 405 NB600-344AF405
Alexa Fluor 488 NB600-344AF488
Alexa Fluor 532 NB600-344AF532
Alexa Fluor 594 NB600-344AF594
Alexa Fluor 647 NB600-344AF647
Alexa Fluor 700 NB600-344AF700
Alexa Fluor 750 NB600-344AF750
Biotin NB600-344B
DyLight 350 NB600-344UV
DyLight 405 NB600-344V
DyLight 488 NB600-344G
DyLight 550 NB600-344R
DyLight 594 NB600-344DL594
DyLight 650 NB600-344C
DyLight 680 NB600-344FR
DyLight 755 NB600-344IR
FITC NB600-344F
HRP NB600-344H
Janelia Fluor 549 NB600-344JF549
Janelia Fluor 646 NB600-344JF646

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Hoist the DYKDDDDK Epitope Tag
The DYKDDDDK Epitope Tag antibody was raised against a short, artificial sequence that is widely and commonly used as a protein fusion tag. It is pervasively present in a vast amount of scientific studies, and is a very powerful investigative research...  Read full blog post.

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