DYKDDDDK Epitope Tag Antibody (29E4.G7) [HRP]

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Product Details

Summary
Reactivity All-NASpecies Glossary
Applications WB, Simple Western, ELISA, IHC, IHC-P
Clone
29E4.G7
Clonality
Monoclonal
Host
Mouse
Conjugate
HRP

Order Details

DYKDDDDK Epitope Tag Antibody (29E4.G7) [HRP] Summary

Immunogen
DYKDDDDK Epitope Tag Antibody (29E4.G7) was produced in mice by repeated immunizations with a synthetic peptide corresponding to the FLAG epitope tag peptide DYKDDDDK (Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys) conjugated to KLH using maleimide. Residues of glycine and cysteine were added to the termini to facilitate coupling.
Specificity
DYKDDDDK Epitope Tag Antibody (29E4.G7) is directed against the motif and is useful in determining its presence in various assays where the epitope tag is present at either the amino or carboxy terminus of recombinant proteins. This monoclonal tag antibody detects over-expressed proteins containing the epitope tag. In Western blotting of bacterial extracts, the antibody does not cross-react with endogenous proteins.
Isotype
IgG2a Kappa
Clonality
Monoclonal
Host
Mouse
Purity
Protein A purified
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Applications/Dilutions

Dilutions
  • Western Blot
  • Simple Western
  • ELISA
  • Immunohistochemistry
  • Immunohistochemistry-Paraffin
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.

Reactivity Notes

Human reactivity reported in a Verified Customer Review.

Packaging, Storage & Formulations

Storage
Store at 4C in the dark.
Buffer
PBS
Preservative
No Preservative
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 (29E4.G7) [HRP]

  • 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

FAQs for DYKDDDDK Epitope Tag Antibody (NBP1-97410H). (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

Other Available Formats

Alexa Fluor 350 NBP1-97410AF350
Alexa Fluor 405 NBP1-97410AF405
Alexa Fluor 488 NBP1-97410AF488
Alexa Fluor 532 NBP1-97410AF532
Alexa Fluor 594 NBP1-97410AF594
Alexa Fluor 647 NBP1-97410AF647
Alexa Fluor 700 NBP1-97410AF700
Alexa Fluor 750 NBP1-97410AF750
Allophycocyanin NBP1-97410APC
Biotin NBP1-97410B
Biotin NBP1-97400
DyLight 350 NBP1-97410UV
DyLight 405 NBP1-97410V
DyLight 488 NBP1-97410G
DyLight 550 NBP1-97410R
DyLight 594 NBP1-97410DL594
DyLight 650 NBP1-97410C
DyLight 680 NBP1-97410FR
DyLight 755 NBP1-97410IR
FITC NBP1-97410F
HRP NBP1-97410H
Janelia Fluor 549 NBP1-97410JF549
Janelia Fluor 646 NBP1-97410JF646
PE NBP1-97410PE
PerCP NBP1-97410PCP

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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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