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Anti-DYKDDDDK Tag Antibody (C-Terminus) LS-C154223

Ordering

Wt. Vol. Conc. Price
250 µg - 1 mg/ml $295
Inquire for larger quantities

LSBio (Direct) LSBio (Direct)
206-374-1102
866-206-6909
Orders@LSBio.com
 

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Details

DYKDDDDK Tag
Rabbit
IgG Polyclonal
Unconjugated
Affinity purified
Unmodified

Applications

  • IHC
  • Western blot (1:2000 - 1:10000)
  • ELISA

Specificity and Use

DYKDDDDK Tag antibody was raised against this antibody was purified from whole rabbit serum prepared by repeated immunizations with the Enterokinase Cleavage Site (ECS) peptide DYKDDDDK (Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys) conjugated to KLH using maleimide. Residues of glycine and cysteine were added to the carboxy terminal end to facilitate coupling. This antibody reacts with FLAG conjugated proteins.
C-Terminus
This affinity purified antibody is directed against DYKDDDDK and is useful in determining its presence in various assays. This polyclonal antibody detects over-expressed proteins containing DYKDDDDK. In western blotting of bacterial extracts, the antibody does not cross-react with endogenous proteins.
This antibody is optimally suited for monitoring the expression of DYKDDDDK tagged fusion proteins. As such, this antibody can be used to identify fusion proteins containing the DYKDDDDK epitope. The antibody recognizes the epitope tag fused to either the amino- or carboxy- termini of targeted proteins. This antibody has been tested by ELISA and western blotting against both the immunizing peptide and DYKDDDDK containing recombinant proteins. Although not tested, this antibody is likely functional for immunoprecipitation, immunocytochemistry, and other immunodetection techniques. The epitope tag peptide sequence was first derived from the 11-amino-acid leader peptide of the gene-10 product from bacteriophage T7. Now the most commonly used hydrophilic octapeptide is DYKDDDDK. This polyclonal antibody to detect DYKDDDDK conjugated proteins binds DYKDDDDK containing fusion proteins with greater affinity than the widely used monoclonal M1, M2 and M5 clones, and shows greater sensitivity in most assays. Affinity purification of the polyclonal antibody results in very low background levels in assays and low cross-reactivity with other cellular proteins.

Packaging

0.02 M potassium phosphate, 0.15 M sodium chloride, pH 7.2, 0.01% sodium azide, sterile filtered
Store at -20°C. Aliquot to avoid freeze/thaw cycles. Store undiluted.
For research use only.

About DYKDDDDK Tag


Western blot

Western blot
Antibody for the detection of FLAG conjugated proteins - Western Blot. Affinity Purified Antibody to detect FLAG conjugated proteins detects both C terminal linked and N terminal linked FLAG tagged recombinant proteins by western blot. This antibody was used at a dilution of 1:2500 to detect 1.0 ug of recombinant protein containing either the FLAG epitope tag linked at the carboxy (C) or the amino (N) terminus of the recombinant protein. A 4-20% gradient gel was used to resolve the protein by SDS-PAGE. The protein was transferred to nitrocellulose using standard methods. After blocking, the membrane was probed with the primary antibody for 1 h at room temperature followed by washes and reaction with a 1:10000 dilution of IRDye 800 conjugated Gt-a-Rabbit IgG (H&L) MX10 (code for 30 min at room temperature. LICORs Odyssey Infrared Imaging System was used to scan and process the image. Other detection systems will yield similar results.

Western blot

Western blot
Antibody for the detection of FLAG conjugated proteins - Western Blot. antibody to detect FLAG conjugated proteins is shown to detect as little as 3 ng of amino-terminal FLAG tagged recombinant protein by western blot. This antibody was used at a 1:1000 dilution to detect 3-fold serial dilutions of amino-terminal FLAG-Bacterial Alkaline Phosphatase (BAP) fusion protein (Sigma P-7582) starting at 1.0 ug of protein as shown in lanes 1-6 respectively. A 4-20% gradient gel was used to separate the protein by SDS-PAGE. The protein was transferred to nitrocellulose using standard methods. After blocking, the membrane was probed with the primary antibody for 1 h at room temperature followed by washes and reaction with a 1:10000 dilution of IRDye 800 conjugated Gt-a-Rabbit IgG (H&L) (code for 30 min at room temperature. LICORs Odyssey Infrared Imaging System was used to scan and process the image. Other detection systems will yield similar results.

Requested From: United States
Date Requested: 12/3/2016

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