DYKDDDDK Epitope Tag Antibody

Western Blot: DYKDDDDK Epitope Tag Antibody [NB600-343] - Samples: 200, 100, or 50 ng of E. coli whole cell lysate expressing a multi-tag fusion protein. Antibodies: Affinity purified, chicken [NB600-343] used for WB at 0.2 ug/ml (1:5,000). Detection: Chemiluminescence with an exposure time of 30 seconds. Observed molecular weight ~49 kDa.

Immunohistochemistry: DYKDDDDK Epitope Tag Antibody [NB600-343] - Drosophila adult optic lobe: Distal medulla cells labeled with DYKDDDDK Epitope Tag Antibody [NB600-343]. Image from verified customer review.

Product Discontinued

DYKDDDDK Epitope Tag Antibody Summary

• Immunogen: To produce DYKDDDDK Epitope Tag Antibody, Chickens 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: Chicken
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • ELISA 1:1000-1:30000 (Primary) 1:100-1:500 (for coating
  • Immunocytochemistry/ Immunofluorescence 1:100-1:400
  • Immunohistochemistry
  • Immunoprecipitation 1-4 ug/ml lysate
  • Western Blot 1:1000-1:30000
• Application Notes: Use in Immunohistochemistry reported in a Verified Customer Review.
• 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

Reactivity with Drosophila reported in a Verified Customer Review.

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.

Review for DYKDDDDK Epitope Tag Antibody (NB600-343) (1)

Average Rating: 4

(Based on 1 review)

We have 1 review tested in 1 species: Drosophila.

reviewed by: Verified Customer IHC Drosophila 07/13/2018

Summary

• Application: Immunohistochemistry
• Sample Tested: Adult Drosophila Brain
• Species: Drosophila
• Lot: A1

FAQs for DYKDDDDK Epitope Tag Antibody (NB600-343). (Showing 1 - 7 of 7 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. Hello. We are planning to promote DYKDDDDK Antibodies in our February 2013 newsletter. Thus, we would like to check toxicity of a few products. Could you let us know whether these are lyophilized form?
   • NB600-343 does not come in lyophilized form.
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.