DYKDDDDK Epitope Tag Antibody (29E4.G7) [CoraFluor™ 1]

• Reactivity: ET
• Applications: WB, ELISA, Flow, IHC
• Clone: 29E4.G7
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: CoraFluor 1

DYKDDDDK Epitope Tag Antibody (29E4.G7) [CoraFluor™ 1] Summary

• Description: CoraFluor(TM) 1 is a high performance terbium-based TR-FRET (Time-Resolved Fluorescence Resonance Energy Transfer) or TRF (Time-Resolved Fluorescence) donor for high throughput assay development. CoraFluor(IM) 1 absorbs UV light at approximately 340 nm, and emits at approximately 490 nm, 545 nm, 585 nm and 620 nm. It is compatible with common acceptor dyes that absorb at the emission wavelengths of CoraFluor(TM) 1. CoraFluor(TM) 1 can be used for the development of robust and scalable TR-FRET binding assays such as target engagement, ternary complex, protein-protein interaction and protein quantification assays.
• Immunogen: This DYKDDDDK Epitope Tag Antibody (29E4.G7) was produced in mice by repeated immunizations with a synthetic peptide corresponding to the FLAG(TM) epitope tag peptide DYKDDDDK Epitope Tag Epitope Tag (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: The purified antibody is directed against the FLAG(TM) 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 anti-FLAG(TM) tag antibody detects over-expressed proteins containing the FLAG(TM) 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

Applications/Dilutions

• Dilutions:
  • ELISA
  • Flow Cytometry
  • Immunohistochemistry
  • Western Blot
• Application Notes: Optimal dilution of this antibody should be experimentally determined.

Packaging, Storage & Formulations

• Storage: Store at 4C in the dark. Do not freeze.
• Buffer: PBS
• Preservative: No Preservative
• Purity: Protein A purified

Notes

CoraFluor (TM) is a trademark of Bio-Techne Corp. Sold for research purposes only under agreement from Massachusetts General Hospital. US patent 2022/0025254

Alternate Names for DYKDDDDK Epitope Tag Antibody (29E4.G7) [CoraFluor™ 1]

• 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.

FAQs for DYKDDDDK Epitope Tag Antibody (NBP1-97410CL1). (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.