DYKDDDDK Epitope Tag Antibody (M2.1) - Chimeric - Azide and BSA Free

Novus Biologicals | Catalog # NBP3-08994

Recombinant Monoclonal Antibody
Novus Biologicals
100% Guarantee

Key Product Details

Species Reactivity

Epitope Tag

Applications

Western Blot, ELISA, Immunocytochemistry/ Immunofluorescence

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Rabbit IgG Kappa Clone # M2.1

Format

Azide and BSA Free
View all DYKDDDDK Epitope Tag Primary Antibodies »

Product Specifications

Immunogen

Interleukin 2 genetically fused to the peptided DYKDDDDK, corresponding to the DDDDK epitope tag.

Clonality

Monoclonal

Host

Rabbit

Isotype

IgG Kappa

Scientific Data Images for DYKDDDDK Epitope Tag Antibody (M2.1) - Chimeric - Azide and BSA Free

ELISA: DYKDDDDK Epitope Tag Antibody (M2.1) - Chimeric - Azide and BSA Free [NBP3-08994]

ELISA: DYKDDDDK Epitope Tag Antibody (M2.1) - Chimeric - Azide and BSA Free [NBP3-08994]

ELISA: DYKDDDDK Epitope Tag Antibody (M2.1) - Chimeric [NBP3-08994] - ELISA using M2.1 and Anti-i-motif DNA [iMab]. Binding curves of the rabbit IgG chimeric version of the Anti-DYKDDDDK Epitope Tag antibody M2.1 (NBP3-08994; blue line) and isotype control (NBP2-52638, anti-fluorescein; orange line) to an ELISA plate coated with Anti-i-motif DNA [iMab] at a concentration of 2.5 ug/ml. A 3-fold serial dilution from 3000 to 0.0169 ng/ml was performed using Anti-i-motif DNA [iMab] antibody. For signal detection, a 1:4000 dilution of a Goat Anti-Rabbit IgG antibody was used.

Applications for DYKDDDDK Epitope Tag Antibody (M2.1) - Chimeric - Azide and BSA Free

Application
Recommended Usage

ELISA

Optimal dilutions of this antibody should be experimentally determined.

Immunocytochemistry/ Immunofluorescence

Optimal dilutions of this antibody should be experimentally determined.

Western Blot

Optimal dilutions of this antibody should be experimentally determined.
Application Notes
This chimeric rabbit antibody was made using the variable domain sequences of the original Mouse IgG1 format, for improved compatibility with existing reagents, assays and techniques.

This antibody recognises he widely used DDDDK-tag commonly used as an epitope tag in fusion proteins. The tag can be recognised at any position within a fusion protein, i.e. N-terminally, Met-N-terminally, C-terminally as well as internally. Binding of this antivbody is not sensitive to calcium. This antibody recognises both native and denatured protein.

Formulation, Preparation, and Storage

Purification

Protein A purified

Formulation

PBS

Format

Azide and BSA Free

Preservative

0.02% Proclin 300

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 (M2.1) - Chimeric - Azide and BSA Free

Certificate of Analysis

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Product Specific Notices for DYKDDDDK Epitope Tag Antibody (M2.1) - Chimeric - Azide and BSA Free

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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Protocols

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

FAQs for DYKDDDDK Epitope Tag Antibody (M2.1) - Chimeric - Azide and BSA Free

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
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