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Anti-DYKDDDDK Tag Antibody (clone 29E4.G7) LS-C154020


Wt. Vol. Conc. Price
- 1000 µl - Unavailable

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Mouse Monoclonal [clone 29E4.G7] (IgG)
Western blot, Immunoprecipitation


IgG Monoclonal [29E4.G7]


  • Western blot
  • Immunoprecipitation

Specificity and Use

DYKDDDDK Tag antibody was raised against this antibody was produced in mice by repeated immunizations with a synthetic peptide corresponding to the FLAG epitope tag peptide DYKDDDDK (Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys) conjugated to KLH.
This antibody is directed against DYKDDDDK and is useful in determining its presence in over expressed proteins in various assays. The antibody recognizes DYKDDDDK fused to either the amino- or carboxy- termini of targeted proteins in transfected or transformed cells.
This agarose-conjugated antibody is optimally suited for immunoprecipitation and purification of DYKDDDDK tagged fusion proteins. The antibody recognizes the epitope tag fused to either the amino- or carboxy- termini of targeted proteins. The epitope tag peptide sequence was first derived from the 11-amino-acid leader peptide of the gene-10 product from bacteriophage T7. DYKDDDDK is the most commonly used hydrophilic octapeptide tag. DYKDDDDK tag can be cleaved by the enterokinase because it contains the Enterokinase Cleavage Site (ECS). DO NOT USE A MAGNETIC STIR BAR WITH THIS PRODUCT. Do not use chaotropic reagents such as guanidine HCl; however urea may be used at less than 1 M concentration. Do not use any reagent that contains reducing agents such as DTT, DTE, or 2-mercaptoethanol with this product. Do not exceed 5% concentration of TWEEN 20 or TRITON X-100 with this product. Do not exceed 0.1% IGEPAL CA-630 or CHAPS. Do not exceed 0.2% digitonin. Do not exceed 1.0 M sodium chloride. DO NOT USE deoxycholate with this product. DO NOT USE SDS. Keep exposure to glycine HCl to less than 20 minutes.


0.02 M potassium phosphate, 0.15 M sodium chloride, pH 7.2, 0.01% sodium azide
Store at 4°C.
For research use only.


Western blot

Western blot
FLAG-Agarose Conjugated - SDS-PAGE. Recombinant FLAG-tagged protein was over-expressed and prepared as a crude E. coli lysate. Approximately 500 ul of the lysate was incubated as slurry with 100 ul of anti-DYKDDDK affinity agarose resin for 1 hour at ambient temperature. The slurry was centrifuged and the flow-through was collected and analyzed by SDS-PAGE. The resin was washed with PBS, 3 x 500 ul. The bound material was then eluted using 0.1 M glycine pH 2.5 and neutralized in 1 M TRIS and also analyzed by SDS-PAGE. Lane 1 shows the crude lysate containing over-expressed DYKDDDK-tagged recombinant protein (approximately 70 kD). Lane 2 shows the unbound flow-through of consisting of endogenous E. coli proteins. Lane 3 shows a prominent band at 70 kD corresponding to enriched recombinant protein.

Western blot

Western blot
FLAG-Agarose Conjugated - SDS-PAGE of Simple Purification. FLAG- Agarose Conjugated Antibody. Briefly, a 500 ul amount of E. coli cell lysate was added to (bind to) 100 ul Anti-FLAG resin for 1 hour at room temperature. After washing 3 x with 750 ul PBS, bound FLAG-protein was eluted in 200 ul of 0.1 M glycine, pH 2.5 followed with neutralization. Cell lysate, flow through and eluate were loaded on SDS gel for quantification. Lanes (6 ul per lane): M) Molecular Weight markers. 1) Cell lysate before purification. 2) Flow through (used cell lysate). 3) Purified FLAG-tagged recombinant protein (arrowhead).

Requested From: 
Date Requested: 4/23/2017

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