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DYKDDDDK Epitope Tag Antibody

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Western Blot: DYKDDDDK Epitope Tag Antibody [NBP2-37833] - Analysis of recombinant protein containing the epitope tag sequence DYKDDDDK using DYKDDDDK Epitope Tag Antibody [NBP2-37833].
Western Blot: DYKDDDDK Epitope Tag Antibody [NBP2-37833] - Analysis of recombinant protein containing the epitope tag sequence DYKDDDDK using DYKDDDDK Epitope Tag Antibody [NBP2-37833].

Product Details

Summary
Reactivity ETSpecies Glossary
Applications WB, ICC/IF
Clonality
Polyclonal
Host
Rabbit
Conjugate
Unconjugated
Concentration
1 mg/ml

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DYKDDDDK Epitope Tag Antibody Summary

Immunogen
DYKDDDDK Epitope Tag Antibody was made to a Synthetic Peptide: D Y K D D D D K C
Specificity
DYKDDDDK Epitope Tag Antibody detects the DYKDDDDK epitope tag sequence.
Isotype
IgG
Clonality
Polyclonal
Host
Rabbit
Purity
Immunogen affinity purified
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Applications/Dilutions

Dilutions
  • Immunofluorescence 1 ug/ml
  • Western Blot 1:500
Application Notes
DYKDDDDK Epitope Tag Antibody has been successfully used in Western blot and immunofluorescence procedures to detect the presence of fusion proteins containing the epitope tag sequence DYKDDDDK.
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.

Packaging, Storage & Formulations

Storage
Store at -20C. Avoid freeze-thaw cycles.
Buffer
PBS with 1 mg/ml BSA
Preservative
0.05% Sodium Azide
Concentration
1 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.

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Product General Protocols

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Video Protocols

WB Video Protocol

FAQs for DYKDDDDK Epitope Tag Antibody (NBP2-37833). (Showing 1 - 6 of 6 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.
  6. What is the immunogen sequence of this FLAG Tag antibody?
    • This antibody was made to a Synthetic Peptide: D Y K D D D D K C

Secondary Antibodies

 

Isotype Controls

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