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Key Product Details

Species Reactivity

Validated:

Epitope Tag

Cited:

Primate

Applications

Validated:

Immunoprecipitation, Western Blot

Cited:

Immunoprecipitation

Label

Agarose

Antibody Source

Polyclonal Goat IgG

Concentration

Please see the vial label for concentration. If unlisted please contact technical services.

Product Specifications

Immunogen

To produce DYKDDDDK Epitope Tag Antibody, Goats were immunized with ECS, Enterokinase Cleavage Site (xxxDDDDK), conjugated to KLH. Antibody was isolated by affinity chromatography using the peptide immobilized on solid support.

Specificity

DYKDDDDK Epitope Tag Antibody reacts with FLAG.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Theoretical MW

1.01 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for DYKDDDDK Epitope Tag Antibody [Agarose]

Western Blot: DYKDDDDK Epitope Tag Antibody [Agarose] [NB600-351] - WB analysis of DYKDDDK in transfected COS-7 cells using [NB600-351]. Image from verified customer review.

Applications for DYKDDDDK Epitope Tag Antibody [Agarose]

Application
Recommended Usage

Western Blot

1:100-1:2000
Application Notes
Final concentration is 0.1mg antibody in 0.4ml of 50% slurry containing 0.2ml agarose. For IP use 15 to 25 ul of gel slurry per 0.1 to 1 mg of protein lysate or extract. Immunoprecipitation was reported in scientific literature (PMID: 23178883) and a Verified Customer Review.

Reviewed Applications

Read 2 reviews rated 4 using NB600-351 in the following applications:

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS

Preservative

0.09% Sodium Azide

Concentration

Please see the vial label for concentration. If unlisted please contact technical services.

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C. Do not freeze.

Background: DYKDDDDK Epitope Tag

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.

Alternate Names

FLAG Tag

Additional DYKDDDDK Epitope Tag Products

Product Documents for DYKDDDDK Epitope Tag Antibody [Agarose]

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Product Specific Notices for DYKDDDDK Epitope Tag Antibody [Agarose]

FLAG(R) and ANTI-FLAG(R) are registered trademarks of Sigma-Aldrich Co. LLC.

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.

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