DYKDDDDK Epitope Tag Antibody (1042E) [Biotin]

R&D Systems | Catalog # BAM8529

Recombinant Monoclonal Antibody.
R&D Systems

Key Product Details

Species Reactivity

Multi-Species

Applications

Western Blot

Label

Biotin

Antibody Source

Recombinant Monoclonal Rabbit IgG Clone # 1042E
View all DYKDDDDK Epitope Tag Primary Antibodies »

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 Antibody (1042E) [Biotin]

Detection of DYKDDDDK (FLAG®) Epitope Tag antibody by Western Blot.

Detection of DYKDDDDK (FLAG®) Epitope Tag by Western Blot.

Western blot shows lysates of HEK293 human embryonic kidney cell line transfected with C-terminal DYKDDDDK-tagged Abro1 and N-terminal DYKDDDDK-tagged RNF20. PVDF membrane was probed with 0.1 µg/mL of Rabbit Anti-DYKDDDDK Biotinylated Monoclonal Antibody (Catalog # BAM8529) followed by Streptavidin-HRP (Catalog # DY998). Specific bands were detected for DYKDDDDK-tagged proteins at approximately 55 and 130 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Applications for DYKDDDDK Epitope Tag Antibody (1042E) [Biotin]

Application
Recommended Usage

Western Blot

0.1 µg/mL
Sample: HEK293 human embryonic kidney cell line transfected with C-terminal DYKDDDDK-tagged Abro1 and N-terminal DYKDDDDK-tagged RNF20

Reviewed Applications

Read 1 review rated 4 using BAM8529 in the following applications:

Formulation, Preparation, and Storage

Purification

Protein A or G purified from cell culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with BSA as a carrier protein.

Shipping

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

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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 (1042E) [Biotin]

Certificate of Analysis

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Product Specific Notices for DYKDDDDK Epitope Tag Antibody (1042E) [Biotin]

FLAG® is a registered trademark of SigmaAldrich Co. LLC

For research use only

Customer Reviews for DYKDDDDK Epitope Tag Antibody (1042E) [Biotin] (1)

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  • DYKDDDDK Epitope Tag Biotinylated Antibody
    Name: Anonymous
    Application: Octet Binding Assay
    Sample Tested: Purified protein
    Species: Human
    Verified Customer | Posted 03/18/2020
    In an octet binding assay, streptavidin biosensors are able to capture the DYKDDDDK epitope tag biotinylated antibody. The DYKDDDDK epitope tag biotinylated antibody can then bind flag tagged proteins.
    DYKDDDDK Epitope Tag Antibody (1042E) [Biotin] BAM8529

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