DYKDDDDK Epitope Tag Antibody (OTI4C5)

Novus Biologicals | Catalog # NBP1-71705

Novus Biologicals
Discontinued Product
NBP1-71705 has been discontinued. View all DYKDDDDK Epitope Tag products.

Key Product Details

Species Reactivity

Non-species specific

Applications

Western Blot, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Simple Western, Immunoprecipitation

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # OTI4C5
View all DYKDDDDK Epitope Tag Primary Antibodies »

Product Specifications

Immunogen

Anti-DDK monoclonal antibody is produced by immunizing mice with a synthetic peptide (DYKDDDDK) coupled to KLH.

Reactivity Notes

DDK epitope tag (DYKDDDDK)

Specificity

The anti-DDK (also known as Flag (R) ) antibody recognizes over expressed recombinant proteins containing the DDK epitope tag (DYKDDDDK) fused to either the amino- or carboxy-termini of targeted proteins in transfected mammalian cells or other expression systems.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

Applications for DYKDDDDK Epitope Tag Antibody (OTI4C5)

Application
Recommended Usage

Flow Cytometry

1:1000

Immunocytochemistry/ Immunofluorescence

1:1000

Immunoprecipitation

1:200

Simple Western

1:250

Western Blot

1:2000
Application Notes

Only recognizes n-terminal form in Simple Western. In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue. This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag (NBP1-71705) present on the protein construct.

Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading.

Flow Cytometry Panel Builder

Bio-Techne Knows Flow Cytometry

Save time and reduce costly mistakes by quickly finding compatible reagents using the Panel Builder Tool.

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
Build Your Panel Now
Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein A or G purified

Formulation

PBS (pH 7.3), 1.0% BSA and 50% Glycerol

Preservative

0.02% 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 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 (OTI4C5)

Certificate of Analysis

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

Download SDS

Product Specific Notices for DYKDDDDK Epitope Tag Antibody (OTI4C5)

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.

Citations for DYKDDDDK Epitope Tag Antibody (OTI4C5)

Customer Reviews for DYKDDDDK Epitope Tag Antibody (OTI4C5)

There are currently no reviews for this product. Be the first to review DYKDDDDK Epitope Tag Antibody (OTI4C5) and earn rewards!

Have you used DYKDDDDK Epitope Tag Antibody (OTI4C5)?

Submit a review and receive an Amazon gift card!

$25/€18/£15/$25CAN/¥2500 Yen for a review with an image

$10/€7/£6/$10CAN/¥1110 Yen for a review without an image

Submit a review
Amazon Gift Card

Protocols

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs for DYKDDDDK Epitope Tag Antibody (OTI4C5)

Showing  1 - 5 of 5 FAQs Showing All

  • Q: Are the FLAG Tag antibodies validated in Simple Western?

    A: Yes, we offer 3 FLAG Tag antibodies that have been tested in Simple Western: NBP1-71705, NBP1-97410, and MAB8529.

  • Q: Do FLAG Tag antibodies comes in lyophilized form?

    A: The following FLAG Tag antibodies come in lyophilized form: NBP1-97393, NBP1-97401, NBP1-97399, NBP1-97400, and NBP1-97402.

  • Q: What the theoretical molecular weight for FLAG Tag antibodies?

    A: The TMW for DYKDDDDK epitope FLAG Tag is approximately 1.01 kDa.

  • Q: Will FLAG Tag antibodies detect internal FLAG tags? My FLAG sequence is sandwiched between two other protein sequences.

    A: If you are applying your sample(s) to SDS-PAGE, under reducing condition and follow by Western Blot, DYKDDDDK epitope tag antibodies should be able to detect the tag region located internally. However, it depends on the property of your chimeric protein and if your tag is in the hydrophobic or hydrophilic region. Although DYKDDDDK is very hydrophilic, it may not be exposed to the surface if embedded deeply inside the protein with a high content of hydrophobic amino acids with or without covalently-linked modifications such as lipid moieties.

  • Q: Will FLAG Tag antibodies recognize FLAG at the C-terminal end?

    A: Yes, FLAG Tag antibodies can recognize FLAG-tag at both the N- or C-terminal.

  • Q: Are the FLAG Tag antibodies validated in Simple Western?

    A: Yes, we offer 3 FLAG Tag antibodies that have been tested in Simple Western: NBP1-71705, NBP1-97410, and MAB8529.

  • Q: Do FLAG Tag antibodies comes in lyophilized form?

    A: The following FLAG Tag antibodies come in lyophilized form: NBP1-97393, NBP1-97401, NBP1-97399, NBP1-97400, and NBP1-97402.

  • Q: What the theoretical molecular weight for FLAG Tag antibodies?

    A: The TMW for DYKDDDDK epitope FLAG Tag is approximately 1.01 kDa.

  • Q: Will FLAG Tag antibodies detect internal FLAG tags? My FLAG sequence is sandwiched between two other protein sequences.

    A: If you are applying your sample(s) to SDS-PAGE, under reducing condition and follow by Western Blot, DYKDDDDK epitope tag antibodies should be able to detect the tag region located internally. However, it depends on the property of your chimeric protein and if your tag is in the hydrophobic or hydrophilic region. Although DYKDDDDK is very hydrophilic, it may not be exposed to the surface if embedded deeply inside the protein with a high content of hydrophobic amino acids with or without covalently-linked modifications such as lipid moieties.

  • Q: Will FLAG Tag antibodies recognize FLAG at the C-terminal end?

    A: Yes, FLAG Tag antibodies can recognize FLAG-tag at both the N- or C-terminal.

  • Q: Are the FLAG Tag antibodies validated in Simple Western?

    A: Yes, we offer 3 FLAG Tag antibodies that have been tested in Simple Western: NBP1-71705, NBP1-97410, and MAB8529.

  • Q: Do FLAG Tag antibodies comes in lyophilized form?

    A: The following FLAG Tag antibodies come in lyophilized form: NBP1-97393, NBP1-97401, NBP1-97399, NBP1-97400, and NBP1-97402.

  • Q: What the theoretical molecular weight for FLAG Tag antibodies?

    A: The TMW for DYKDDDDK epitope FLAG Tag is approximately 1.01 kDa.

  • Q: Will FLAG Tag antibodies detect internal FLAG tags? My FLAG sequence is sandwiched between two other protein sequences.

    A: If you are applying your sample(s) to SDS-PAGE, under reducing condition and follow by Western Blot, DYKDDDDK epitope tag antibodies should be able to detect the tag region located internally. However, it depends on the property of your chimeric protein and if your tag is in the hydrophobic or hydrophilic region. Although DYKDDDDK is very hydrophilic, it may not be exposed to the surface if embedded deeply inside the protein with a high content of hydrophobic amino acids with or without covalently-linked modifications such as lipid moieties.

  • Q: Will FLAG Tag antibodies recognize FLAG at the C-terminal end?

    A: Yes, FLAG Tag antibodies can recognize FLAG-tag at both the N- or C-terminal.

  • Q: Are the FLAG Tag antibodies validated in Simple Western?

    A: Yes, we offer 3 FLAG Tag antibodies that have been tested in Simple Western: NBP1-71705, NBP1-97410, and MAB8529.

  • Q: Do FLAG Tag antibodies comes in lyophilized form?

    A: The following FLAG Tag antibodies come in lyophilized form: NBP1-97393, NBP1-97401, NBP1-97399, NBP1-97400, and NBP1-97402.

  • Q: What the theoretical molecular weight for FLAG Tag antibodies?

    A: The TMW for DYKDDDDK epitope FLAG Tag is approximately 1.01 kDa.

  • Q: Will FLAG Tag antibodies detect internal FLAG tags? My FLAG sequence is sandwiched between two other protein sequences.

    A: If you are applying your sample(s) to SDS-PAGE, under reducing condition and follow by Western Blot, DYKDDDDK epitope tag antibodies should be able to detect the tag region located internally. However, it depends on the property of your chimeric protein and if your tag is in the hydrophobic or hydrophilic region. Although DYKDDDDK is very hydrophilic, it may not be exposed to the surface if embedded deeply inside the protein with a high content of hydrophobic amino acids with or without covalently-linked modifications such as lipid moieties.

  • Q: Will FLAG Tag antibodies recognize FLAG at the C-terminal end?

    A: Yes, FLAG Tag antibodies can recognize FLAG-tag at both the N- or C-terminal.

  • Q: Are the FLAG Tag antibodies validated in Simple Western?

    A: Yes, we offer 3 FLAG Tag antibodies that have been tested in Simple Western: NBP1-71705, NBP1-97410, and MAB8529.

  • Q: Do FLAG Tag antibodies comes in lyophilized form?

    A: The following FLAG Tag antibodies come in lyophilized form: NBP1-97393, NBP1-97401, NBP1-97399, NBP1-97400, and NBP1-97402.

  • Q: What the theoretical molecular weight for FLAG Tag antibodies?

    A: The TMW for DYKDDDDK epitope FLAG Tag is approximately 1.01 kDa.

  • Q: Will FLAG Tag antibodies detect internal FLAG tags? My FLAG sequence is sandwiched between two other protein sequences.

    A: If you are applying your sample(s) to SDS-PAGE, under reducing condition and follow by Western Blot, DYKDDDDK epitope tag antibodies should be able to detect the tag region located internally. However, it depends on the property of your chimeric protein and if your tag is in the hydrophobic or hydrophilic region. Although DYKDDDDK is very hydrophilic, it may not be exposed to the surface if embedded deeply inside the protein with a high content of hydrophobic amino acids with or without covalently-linked modifications such as lipid moieties.

  • Q: Will FLAG Tag antibodies recognize FLAG at the C-terminal end?

    A: Yes, FLAG Tag antibodies can recognize FLAG-tag at both the N- or C-terminal.

Showing  1 - 5 of 5 FAQs Showing All
View all FAQs for Antibodies