Key Product Details

Validated by

Biological Validation

Species Reactivity

Epitope Tag

Applications

Immunocytochemistry/ Immunofluorescence, Immunoprecipitation, Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2B Clone # FG4R

Format

Azide and BSA Free

Concentration

1 mg/ml

View all DYKDDDDK Epitope Tag Antibodies »

Product Specifications

Immunogen

DYKDDDDK Epitope Tag Antibody (FG4R) was made to a Synthetic peptide (DYKDDDDK) coupled to KLH.

Specificity

DYKDDDDK Epitope Tag Antibody (FG4R) detects Flag tag from various samples.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2B

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 (FG4R) - Azide and BSA Free

Western Blot: DYKDDDDK Epitope Tag Antibody (FG4R)Azide and BSA Free [NBP2-37823]

Western Blot: DYKDDDDK Epitope Tag Antibody (FG4R)Azide and BSA Free [NBP2-37823]

Western Blot: DYKDDDDK Epitope Tag Antibody (FG4R) [NBP2-37823] - Analysis using [NBP2-37823] at a 1:1000 (1ug/mL) dilution, probed against HEK293 cells transfected with DYKDDDDK-tagged protein vector; untransfected (1) and transfected (2). Observed molecular weight ~130 kDa.
Immunocytochemistry/ Immunofluorescence: DYKDDDDK Epitope Tag Antibody (FG4R) - Azide and BSA Free [NBP2-37823]

Immunocytochemistry/ Immunofluorescence: DYKDDDDK Epitope Tag Antibody (FG4R) - Azide and BSA Free [NBP2-37823]

Immunocytochemistry/Immunofluorescence: DYKDDDDK Epitope Tag Antibody (FG4R) [NBP2-37823] - Analysis of Chlamydia trachomatis FLAG-tagged IncA (blue) in HeLa cells. Cells were infected with a GFP-expressing strain of Chlamydia trachomatis containing FLAG-IncA under the control of the Tetracycline (Tet) promoter, and either left untreated (0ng/ml Tet) or treated with anhydrotetracycline (10ng/ml Tet). Cells were fixed with 2% paraformaldehyde in PBS for 30 min at RT, permeabilized and blocked in 10% goat serum, 0.1% BSA, 0.1% saponin in PBS pH 7.4 for 30 mins at RT, then probed with a DYKDDDDK Epitope Tag Antibody (FG4R) [NBP2-37823] at 1:500 for 1 hr at RT. Cells were washed with PBS, and incubated with an AlexaFluor 647-conjugated goat anti-mouse IgG secondary antibody at 1:2000 for 1 hr at RT. Images were taken on a fluorescent microscope using a 60X oil-immersion objective. Scale bars represent 20um. Data courtesy of the Innovators Program.
Western Blot: DYKDDDDK Epitope Tag Antibody (FG4R)Azide and BSA Free [NBP2-37823]

Western Blot: DYKDDDDK Epitope Tag Antibody (FG4R)Azide and BSA Free [NBP2-37823]

Western Blot: DYKDDDDK Epitope Tag Antibody (FG4R) [NBP2-37823] - Analysis of 30ug of whole cell lysates from HeLa cells infected with a strain of Chlamydia trachomatis expressing FLAG-IncA under the control of a Tetracycline promoter (Tet), and either left untreated (-) or treated with anhydrotetracycline (+), onto an SDS-PAGE gel, using [NBP2-37823]. Observed molecular weight ~30 kDa.
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Applications for DYKDDDDK Epitope Tag Antibody (FG4R) - Azide and BSA Free

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

1:200 - 1:500

Immunoprecipitation

3 ug

Western Blot

1:500 - 1:1000

Formulation, Preparation, and Storage

Purification

Protein A purified

Formulation

PBS (pH 7.2)

Format

Azide and BSA Free

Preservative

No Preservative

Concentration

1 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 short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

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 (FG4R) - Azide and BSA Free

Product Datasheets

Certificate of Analysis

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Product Specific Notices for DYKDDDDK Epitope Tag Antibody (FG4R) - Azide and 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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