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3X (DYKDDDDK) Peptide

Catalog No.
A6001
3X FLAG peptide
Grouped product items
SizePriceStock Qty
4mg
$90.00
In stock
25mg
$405.00
In stock
For scientific research use only and should not be used for diagnostic or medical purposes.

Tel: +1-832-696-8203

Email: [email protected]

Worldwide Distributors

Background

The 3X FLAG peptide, consisting of three tandem repeats of the sequence DYKDDDDK, is widely utilized in biological research as an epitope tag for recombinant protein detection and purification. Its hydrophilic, 23-residue structure facilitates effective exposure and recognition by monoclonal antibody anti-FLAG M1 or M2(MA5839), improving sensitivity for fusion protein identification. After fusion to target proteins, this peptide tag allows affinity-based isolation or immunodetection methods in various experimental conditions. Due to its small size and hydrophilicity, the 3X FLAG peptide minimally interferes with protein structure and function, including protein crystallization procedures for structural analysis.

References:
1. Hopp TP, Prickett KS, Price VL, Libby RT, March CJ, Ceretti DP, Urdal DL, Conlon PJ (1988) A short polypeptide marker sequence useful for recombinant protein identification and purification. Bio/Technology 6:1204–1210.
2. Hopp TP, Gallis B, Prikett KS (1996) Metal-binding properties of a calcium dependent monoclonal antibody. Mol Immunol 33:601–608.
3. Einhauer A, Jungbauer A (2000) Kinetics and thermodynamical properties of the monoclonal antibody M1 directed against the FLAG peptide. 20th International symposium on the separation of proteins, peptides, and polynucleotides (ISPPP). Lublijana, Slovenia, November 5–8, 2000.
4. Bucher MH, Evdokimov AG, Waugh DS (2002) Differential effects of short affinity tags on the crystallization of Pyrococcus furiosus maltodextrin-binding protein. Biol Cryst 58:392–397.
5. Maroux S, Baratti J, Desnuelle P (1971) Purification and specificity of procine enterokinase. J Biol Chem 246:5031–5039.

Product Citation

Chemical Properties

StorageDesiccate at -20°C
M.Wt2861.87
Cas No.402750-12-3
FormulaC120H169N31O49S
SynonymsH-Met-Asp-Tyr-Lys-Asp-His-Asp-Gly-Asp-Tyr-Lys-Asp-His-Asp-Ile-Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys-OH
Solubility≥25mg/ml in TBS (0.5M Tris-HCl, pH 7.4, with 1M NaCl), 1X TBS as recommended buffer.
Chemical Name3X (DYKDDDDK) Peptide
SDFDownload SDF
Canonical SMILESCCC(C)C(C(=O)NC(CC(=O)O)C(=O)NC(CC1=CC=C(C=C1)O)C(=O)NC(CCCCN)C(=O)NC(CC(=O)O)C(=O)NC(CC(=O)O)C(=O)NC(CC(=O)O)C(=O)NC(CC(=O)O)C(=O)NC(CCCCN)C(=O)O)NC(=O)C(CC(=O)O)NC(=O)C(CC2=CNC=N2)NC(=O)C(CC(=O)O)NC(=O)C(CCCCN)NC(=O)C(CC3=CC=C(C=C3)O)NC(=O)C(CC(=O)O)NC(
Shipping ConditionSmall Molecules with Blue Ice, Modified Nucleotides with Dry Ice.
General tips We do not recommend long-term storage for the solution, please use it up soon.

Protocol

ELISA experiment [1]:

Preparation method

The solubility of this peptide in sterile water is >10 mM. Stock solution should be splited and stored at -80°C for several months.

Applications

3-DYKDDDDK peptide has found widespread use as a mild purification reagent for DYKDDDDK-epitope tagged recombinant proteins. Although its affinity columns release monovalent flagged proteins in the absence of calcium, the antibody retains substantial affinity for the DYKDDDDK sequence even in metal-free conditions, so that it has been impossible to use it to develop a metal-sensitive ELISA assay. This is due to the ability of the antibody to remain bound to polyvalent surface-coated antigen, for instance, when Flagged proteins are bound to ELISA plates or blotting filters. The resultant antigen polyvalence raises the avidity of the DYKDDDDK antibody to a point where the reaction is essentially calcium-independent. However, when the antibody itself was made monovalent, by proteolytic cleavage to the Fab, this situation was reversed and the ELISA reaction became calcium-dependent. This new metal-dependent ELISA assay was used to explore the metal requirements of the antibody in detail. Among divalent metals, binding tapered off with increasing radius above that of calcium, or with decreasing radius below that of calcium. Several smaller metals, such as nickel, acted as inhibitors of the binding reaction. Substantial binding was demonstrated for heavy metals such as cadmium, lanthanum and samarium. Because it is of interest to use this antibody for the co-crystallization of recombinant DYKDDDDK-fusion proteins, the ability to bind heavy metals was a significant finding.

References:

1. Hopp TP1, Gallis B, Prickett KS. Metal-binding properties of a calcium-dependent monoclonal antibody. Mol Immunol. 1996 May-Jun;33(7-8):601-8.

Biological Activity

Description 3X (DYKDDDDK) Peptide is a synthetic peptide with a 3-time repeated DYKXXD motif.
Targets anti-DYKDDDDK M2 antibody          
IC50            

Quality Control

Quality Control & MSDS

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