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Key Product Details

Species Reactivity

Epitope Tag

Applications

ELISA, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG

Format

BSA Free

Concentration

1.0 mg/ml

Product Summary for DYKDDDDK Epitope Tag Antibody - BSA Free

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 - BSA Free

Western Blot: DYKDDDDK Epitope Tag Antibody [NB600-344] - Samples: 200, 100, or 50 ng of E. coli whole cell lysate expressing a multi-tag fusion protein. Antibody: Affinity purified goat DYKDDDDK Epitope Tag Antibody [NB600-344] used for WB at 0.04 ug/ml (1:25,000). Detection: Chemiluminescence with an exposure time of 30 seconds. Observed molecular weight ~49 kDa.
Western Blot: DYKDDDDK Epitope Tag Antibody [NB600-344] - Analysis using the Agarose Immobilized conjugate of DYKDDDDK Epitope Tag Antibody [NB600-344]. DYKDDDK in transfected COS-7 cells using agarose immobilized DYKDDDDK Epitope Tag Antibody.
Immunocytochemistry/Immunofluorescence: DYKDDDDK Epitope Tag Antibody [NB600-344] - Expression of the ZFN as detected by PLA.PK-15 cells were transfected with the pZFN1 or pZFN2 vectors expressing the 3xFLAG tagged fluorescent proteins ZFN1-CFP or ZFN2-YFP, the nucleus was stained by DAPI and a PLA against the FLAG epitope (DYKDDDDK Epitope Tag Antibody [NB600-344]) was performed demonstrating the specific localization of the ZFN in the nucleus. Scale bars, 5 um. Image collected and cropped by CiteAb from the following publication (http://dx.plos.org/10.1371/journal.pone.0122059), licensed under a CC-BY license.

Applications for DYKDDDDK Epitope Tag Antibody - BSA Free

Application
Recommended Usage

ELISA

1:100-1:500 (coating); 1:1000-1:30000 (primary)

Immunocytochemistry/ Immunofluorescence

1:100-1:400

Immunoprecipitation

1-4 ug/mg Lysate

Western Blot

1:1000-1:30000
Application Notes
Use in IP, WB, and ICC/IF reported in multiple pieces of scientific literature.
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Published Applications

Read 25 publications using NB600-344 in the following applications:

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS

Format

BSA Free

Preservative

0.09% Sodium Azide

Concentration

1.0 mg/ml

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

Product Documents for DYKDDDDK Epitope Tag Antibody - BSA Free

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 - BSA Free

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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FAQs for DYKDDDDK Epitope Tag Antibody - BSA Free

Showing  1 - 2 of 2 FAQs Showing All
  • Q: Is this antibody suitable for ChIP analysis of an N-terminally tagged protein?

    A: Our DYKDDDDK epitope tag antibody with catalogue number NB600-344 has been validated for Western blotting, immunofluorescence, immunoprecipitation and ELISA but has not yet been tested for ChIP. However, our DYKDDDDK Epitope Tag antibody with catalog number NBP2-59146 is validated for use in ChIP.

  • Q: What is the immunogen sequence of this FLAG Tag antibody? 

    A: This antibody was made to a ECS enterokinase cleavage site (xxxDDDDK) conjugated to KLH.  

Showing  1 - 2 of 2 FAQs Showing All
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