Key Product Details

Species Reactivity

Validated:

Multi-Species

Cited:

Human, Mouse

Applications

Validated:

Intracellular Staining by Flow Cytometry

Cited:

Flow Cytometry

Label

Alexa Fluor 488 (Excitation = 488 nm, Emission = 515-545 nm)

Antibody Source

Recombinant Monoclonal Rabbit IgG Clone # 1042E
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Product Specifications

Immunogen

DYKDDDDK epitope tag

Specificity

Detects N-terminal and C-terminal DYKDDDDK (FLAG®) epitope tagged proteins in Western blot.

Clonality

Monoclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for DYKDDDDK Epitope Tag Alexa Fluor® 488‑conjugated Antibody

Detection of DYKDDDDK Epitope Tag in HEK293 cells by Flow Cytometry.

HEK293 cells transfected with DYKDDDDK-tagged proteins (filled histogram) vs Irrelevant HEK293 wild type cells (open histogram) were stained with Rabbit Anti-DYKDDDDK Epitope Tag Alexa Fluor® 488‑conjugated Monoclonal Antibody (Catalog # IC8529G) at 0.001 ug/106 cells. To facilitate intracellular staining, cells were fixed and permeabilized with Flow Cytometry Fixation Buffer (Catalog # FC004). View our protocol for Staining Intracellular Molecules.

Applications for DYKDDDDK Epitope Tag Alexa Fluor® 488‑conjugated Antibody

Application
Recommended Usage

Intracellular Staining by Flow Cytometry

Titration recommended for optimal concentration with starting range of 0.001-1 ug/106 cells. Sample used for this experiment was HEK293 cells transfected with FLAG, fixed and permeabilized with Flow Cytometry Fixation Buffer (Catalog # FC004).

Reviewed Applications

Read 1 review rated 4 using IC8529G in the following applications:

Spectra Viewer

Plan Your Experiments

Use our spectra viewer to interactively plan your experiments, assessing multiplexing options. View the excitation and emission spectra for our fluorescent dye range and other commonly used dyes.

Spectra Viewer

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Advanced Features

  • Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
  • Spillover Popups - Visualize the spectra of individual fluorochromes
  • Antigen Density Selector - Match fluorochrome brightness with antigen density
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Formulation, Preparation, and Storage

Purification

Protein A or G purified from cell culture supernatant

Formulation

Supplied in a saline solution containing BSA and Sodium Azide.

Shipping

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

Stability & Storage

Protect from light. Do not freeze.
  • 12 months from date of receipt, 2 to 8 °C as supplied.

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 Alexa Fluor® 488‑conjugated Antibody

Certificate of Analysis

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

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for DYKDDDDK Epitope Tag Alexa Fluor® 488‑conjugated Antibody

FLAG® is a registered trademark of Sigma-Aldrich Co. LLC


This product is provided under an agreement between Life Technologies Corporation and R&D Systems, Inc, and the manufacture, use, sale or import of this product is subject to one or more US patents and corresponding non-US equivalents, owned by Life Technologies Corporation and its affiliates. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The sale of this product is expressly conditioned on the buyer not using the product or its components (1) in manufacturing; (2) to provide a service, information, or data to an unaffiliated third party for payment; (3) for therapeutic, diagnostic or prophylactic purposes; (4) to resell, sell, or otherwise transfer this product or its components to any third party, or for any other commercial purpose. Life Technologies Corporation will not assert a claim against the buyer of the infringement of the above patents based on the manufacture, use or sale of a commercial product developed in research by the buyer in which this product or its components was employed, provided that neither this product nor any of its components was used in the manufacture of such product. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, Cell Analysis Business Unit, Business Development, 29851 Willow Creek Road, Eugene, OR 97402, Tel: (541) 465-8300. Fax: (541) 335-0354.

For research use only

Citations for DYKDDDDK Epitope Tag Alexa Fluor® 488‑conjugated Antibody

Customer Reviews (1)

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  • Name: Anonymous
    Application: Flow Cytometry
    Sample Tested: HEK293 human embryonic kidney cell line
    Species: Human
    Verified Customer | Posted 04/13/2023
    DYKDDDDK Epitope Tag Alexa Fluor® 488‑conjugated Antibody IC8529G

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