Key Product Details

Species Reactivity

Validated:

Epitope Tag

Applications

Validated:

ELISA, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunoprecipitation, Simple Western, Western Blot

Cited:

IF/IHC, IP, SDS-Page, WB

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free

Concentration

1.0 mg/ml

View all DYKDDDDK Epitope Tag Antibodies »

Product Summary for DYKDDDDK Epitope Tag Antibody - BSA Free

Immunogen

To produce DYKDDDDK Epitope Tag Antibody, Rabbits 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

Rabbit

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 [NB600-345] - used for Western Blot. Lane 1: Untransfected HEK (control) Lane 2: Transfected HEK (FLAG-tagged protein, clone 1) Lane 3: Transfected HEK (FLAG-tagged protein, clone 2). Image from verified customer review.
Immunocytochemistry/Immunofluorescence: DYKDDDDK Epitope Tag Antibody [NB600-345] - Samples: Human microvascular endothelial cells expressing FLAG tagged beta-catenin following transient transfection. Antibody: Affinity purified goat DYKDDDDK Epitope Tag Antibody [NB600-345] used at 1 ug/ml. Detection: FITC labeled rabbit anti-goat IgG (h+l).
Western Blot: DYKDDDDK Epitope Tag Antibody [NB600-345] - 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 [NB600-345] 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

Applications for DYKDDDDK Epitope Tag Antibody - BSA Free

Application
Recommended Usage

ELISA

1:1000-1:30000 (Primary) 1:100-1:500 (For Coating

Immunocytochemistry/ Immunofluorescence

1:100-1:400

Immunoprecipitation

1 - 4 ug/mg lysate

Simple Western

reported by internal validation

Western Blot

1:1000-1:30000
Application Notes
Use in IHC reported in scientific literature (PMID: 33219296) ELISA for coating plates 1:100 - 1:500. Use in Western Blot reported in scientific literature (PMID: 30297740). In Simple Western internal validation: Abro1 NT Tx lysate at 0.1 mg/ml as sample; separated by size; antibody dilution of 1:250; matrix was 12-230 kDa; detected by Chemiluminescence.
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 4 reviews rated 4.5 using NB600-345 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

Additional DYKDDDDK Epitope Tag Products

Product Documents for DYKDDDDK Epitope Tag Antibody - BSA Free

Product Datasheets

Certificate of Analysis

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

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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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