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DYKDDDDK Epitope Tag Antibody (F-tag-01) [Biotin]

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Product Details

Summary
Reactivity ETSpecies Glossary
Applications WB, ICC/IF
Clone
F-tag-01
Clonality
Monoclonal
Host
Mouse
Conjugate
Biotin

Order Details

View Available Conjugates
Catalog# & Conjugate Size Price
Alexa Fluor 350
NB500-554AF350
Alexa Fluor 405
NB500-554AF405
Alexa Fluor 488
NB500-554AF488
Alexa Fluor 532
NB500-554AF532
Alexa Fluor 594
NB500-554AF594
Alexa Fluor 647
NB500-554AF647
Alexa Fluor 700
NB500-554AF700
Alexa Fluor 750
NB500-554AF750
CoraFluor 1
NB500-554CL1
DyLight 350
NB500-554UV
DyLight 405
NB500-554V
DyLight 488
NB500-554G
DyLight 550
NB500-554R
DyLight 594
NB500-554DL594
DyLight 650
NB500-554C
DyLight 680
NB500-554FR
DyLight 755
NB500-554IR
FITC
NB500-554F
HRP
NB500-554H
Janelia Fluor 525
NB500-554JF525
Janelia Fluor 549
NB500-554JF549
Janelia Fluor 585
NB500-554JF585
Janelia Fluor 635
NB500-554JF635
Janelia Fluor 646
NB500-554JF646
Janelia Fluor 669
NB500-554JF669
Unconjugated
NB500-554
mFluor Violet 450 SE
NB500-554MFV450
mFluor Violet 500 SE
NB500-554MFV500
mFluor Violet 610 SE
NB500-554MFV610
View Available Formulations
Catalog# & Formulation Size Price
Standard Formulation
NB500-554
BSA Free
NB500-554
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DYKDDDDK Epitope Tag Antibody (F-tag-01) [Biotin] Summary

Immunogen
DYKDDDDK Epitope Tag Antibody (F-tag-01) was made to a Synthetic peptide: DYKDDDDK conjugated to KLH.
Specificity
DYKDDDDK Epitope Tag Antibody (F-tag-01) recognizes DDDDK-tagged proteins in all species. The small size of this tag and its high hydrophilicity decrease the probability of interference with its expression, proteolytic maturation, antigenicity, localization and function.
Isotype
IgG1
Clonality
Monoclonal
Host
Mouse
Purity
Protein A purified
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Applications/Dilutions

Dilutions
  • Immunocytochemistry/ Immunofluorescence
  • Western Blot
Application Notes
Optimal dilution of this antibody should be experimentally determined.
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.
Readout System

Reactivity Notes

Tag specific

Packaging, Storage & Formulations

Storage
Store at 4C in the dark.
Buffer
PBS
Preservative
0.05% Sodium Azide
Purity
Protein A purified

Notes

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

Alternate Names for DYKDDDDK Epitope Tag Antibody (F-tag-01) [Biotin]

  • 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 (NB500-554B). (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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