DYKDDDDK Epitope Tag Antibody (L5) [DyLight 488]

Immunocytochemistry/Immunofluorescence: Rat Monoclonal DYKDDDDK Epitope Tag Antibody (L5) [DyLight 488] [NBP1-06712G] - Normal mouse prostate cells overexpressed with Flag-tagged construct (Green: Flag). Image from verified customer review.

• Reactivity: ET
• Applications: WB, ICC/IF, IHC, IP
• Clone: L5
• Clonality: Monoclonal
• Host: Rat
• Conjugate: DyLight 488

DYKDDDDK Epitope Tag Antibody (L5) [DyLight 488] Summary

• Description: This conjugate is made on demand and is not held in inventory. Each order represents a unique lot. All conjugates are generated using validated conjugation protocols and meet strict degree of labeling (F/P) specifications; additional information is available upon request. The antibody concentration is provided on the individual product label.
  
  For specialized conjugation needs, like large quantities, specific concentrations, and defined F/P ratios, bulk and custom antibody conjugation services are available.
• Immunogen: DYKDDDDK Epitope Tag Antibody (L5) was made to N-terminal DYKDDDDK-tagged extracellular domain of mouse Langerin. Binds to same epitope as Sigma's Anti-FLAG® M2 Antibody.
• Epitope: DDDDK
• Specificity: Binds to same epitope as Sigma's Anti-FLAGM2 Antibody. FLAG is a registered trademark of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co.
• Isotype: IgG2a
• Clonality: Monoclonal
• Host: Rat
• Purity: Tissue culture supernatant

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry/ Immunofluorescence
  • Immunohistochemistry
  • Immunohistochemistry-Frozen
  • Immunoprecipitation
  • Western Blot
• Application Notes: Recommended applications are based on validated applications from the unconjugated base product NBP1-06712. This conjugated antibody is not kept in inventory and is made to order using validated conjugation protocols.

Packaging, Storage & Formulations

• Storage: Store at 4C in the dark.
• Buffer: 50mM Sodium Borate
• Preservative: 0.05% Sodium Azide
• Purity: Tissue culture supernatant

Notes

DyLight (R) is a trademark of Thermo Fisher Scientific Inc. and its subsidiaries.

Alternate Names for DYKDDDDK Epitope Tag Antibody (L5) [DyLight 488]

• 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 (NBP1-06712G) (1)

Average Rating: 4

(Based on 1 review)

We have 1 review tested in 1 species: Mouse.

reviewed by: Xuanrong Chen ICC Mouse 09/01/2023

Summary

• Application: Immunocytochemistry
• Sample Tested: Normal mouse prostate cells
• Species: Mouse
• Lot: D145957

FAQs for DYKDDDDK Epitope Tag Antibody (NBP1-06712G). (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.