DYKDDDDK tag Polyclonal antibody (Binds to FLAG® tag epitope)

DYKDDDDK tag Polyclonal Antibody for IF, WB, ELISA

Host / Isotype

Rabbit / IgG

Reactivity

recombinant protein and More (2)

Applications

WB, RIP, IP, IHC, IF, FC, CoIP, ELISA

Conjugate

Unconjugated

Cat no : 20543-1-AP

Synonyms

DDDDK tag, DYKDDDDK, Flag



Tested Applications

Positive WB detected inrecombinant protein protein
Positive IF detected inTransfected HEK-293 cells

Recommended dilution

ApplicationDilution
Western Blot (WB)WB : 1:200-1:1000
Immunofluorescence (IF)IF : 1:10-1:100
Sample-dependent, check data in validation data gallery

Product Information

The immunogen of 20543-1-AP is DYKDDDDK tag Fusion Protein expressed in E. coli.

Tested Reactivity recombinant protein
Cited Reactivity human, mouse, recombinant protein
Host / Isotype Rabbit / IgG
Class Polyclonal
Type Antibody
Immunogen DYKDDDDK tag fusion protein Ag2329
Full Name DYKDDDDK tag
Gene symbol
Gene ID (NCBI)
Conjugate Unconjugated
Form Liquid
Purification Method Antigen affinity purification
Storage Buffer PBS with 0.1% sodium azide and 50% glycerol pH 7.3.
Storage ConditionsStore at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20oC storage.

Background Information

Protein tags are protein or peptide sequences located either on the C- or N- terminal of the target protein, which facilitates one or several of the following characteristics: solubility, detection, purification, localization and expression. The DYKDDDDK(FLAG) peptide has been used extensively as a general tag in expression vectors. This peptide can be expressed and detected with the protein of interest as an amino-terminal or carboxy-terminal fusion. N-terminal DDDDK vectors provide an Ek cleavage site for removal of the fusion tag. The DDDDK peptide is likely to be located on the surface of a fusion protein because of its hydrophilic nature. As a result, the DDDDK peptide is more likely to be accessible to antibodies. A DDDDK-tag can be used in many different assays that require recognition by an antibody, such as western blotting, immunocytochemistry, immunoprecipitation, flow cytometry, protein purification, and in the study of protein-protein interactions, cell ultrastructure, and protein localization and so on. This antibody is a rabbit polyclonal antibody raised against 3xFlag (3xDYKDDDDKT) sequence and recognizes the (1x) and (3x)DYKDDDDK peptide and detects DDDDK-tagged proteins. Anti-FLAG is a registered trademark of Sigma-Aldrich Biotechnology.

Protocols

Product Specific Protocols
WB protocol for DYKDDDDK tag antibody 20543-1-APDownload protocol
IF protocol for DYKDDDDK tag antibody 20543-1-APDownload protocol
Standard Protocols
Click here to view our Standard Protocols

Publications

SpeciesApplicationTitle

Front Immunol

Long Interleukin-22 Binding Protein Isoform-1 Is an Intracellular Activator of the Unfolded Protein Response.

Authors - Paloma Gómez-Fernández

Mol Immunol

CpG oligodeoxynucleotide-specific duck TLR21 mediates activation of NF-κB signaling pathway and plays an important role in the host defence of DPV infection.

Authors - Da Cheng

Viruses

PSMB1 Negatively Regulates the Innate Antiviral Immunity by Facilitating Degradation of IKK-ε.

Authors - Fangyi Wu

Cell Death Differ

Glutathione S-transferases P1 protects breast cancer cell from adriamycin-induced cell death through promoting autophagy.

Authors - Xiaoliang Dong

Biochem Pharmacol

PMID: 30689982

Authors - Min Chen

Free Radic Biol Med

SIRT5-mediated SDHA desuccinylation promotes clear cell renal cell carcinoma tumorigenesis.

Authors - Yuanzhen Ma

Reviews

The reviews below have been submitted by verified Proteintech customers who received an incentive forproviding their feedback.


FH

Tongbin (Verified Customer) (08-25-2020)

For western blots, this FLAG antibody is comparable to Sigma's M2 FLAG antibody, which has been the gold standard for FLAG.

  • Applications: Western Blot
  • Primary Antibody Dilution: 1:1000
  • Cell Tissue Type: HEK293T
FH

Dipen (Verified Customer) (11-11-2019)

Great antibody for Western blotting. I used it at 1:1000 and got a super strong signal. Will use at a lower concentration next time.

  • Applications: Western Blot,
  • Primary Antibody Dilution: 1:1000
  • Cell Tissue Type: HEK293T cells
DYKDDDDK tag Antibody Western Blot, validation (1:1000 dilution) in HEK293T cells (Cat no:20543-1-AP)
FH

Sara (Verified Customer) (10-24-2019)

Works really well for both IP of overexpressed FLAG-tagged proteins and WB!

  • Applications: Western Blot,Immunoprecipitation,
  • Primary Antibody Dilution: 1:2000
  • Cell Tissue Type: 293T overexpressing FLAG-tagged construct
DYKDDDDK tag Antibody Western Blot,Immunoprecipitation, validation (1:2000 dilution) in 293T overexpressing FLAG-tagged construct (Cat no:20543-1-AP)
FH

Nikhil (Verified Customer) (10-16-2019)

Flag tagged p53 expressed in H1299 cells.

  • Applications: Western Blot,
  • Primary Antibody Dilution: 1:1000
  • Cell Tissue Type: H1299
DYKDDDDK tag Antibody Western Blot, validation (1:1000 dilution) in H1299 (Cat no:20543-1-AP)
FH

Shivani (Verified Customer) (09-26-2019)

Works better than another brand I had tested.

  • Applications: Western Blot,
  • Primary Antibody Dilution: 1:1000
FH

Corey (Verified Customer) (08-23-2019)

This antibody has worked well for western blotting as well as immunoprecipitation. The lots have been consistent so far, we will definitely be ordering more.

  • Applications: Western Blot,Immunoprecipitation,
  • Primary Antibody Dilution: 1:1000
  • Cell Tissue Type: H1299
FH

Kishor (Verified Customer) (06-23-2019)

Works very good for WB at 1:1000 and IP at 1:200.

  • Applications: Western Blot,Immunoprecipitation,
  • Primary Antibody Dilution: 1:1000
  • Cell Tissue Type: Human cell lines
FH

David (Verified Customer) (06-19-2019)

This is working very well for me. I ran out of another brand and got this one because I saw you had it and it was a better value. After testing it in the lab, it works just as well, if not better than the previous one. A very strong signal in my experiments.

  • Applications: Western Blot,
  • Primary Antibody Dilution: 1/2000
FH

Lana (Verified Customer) (04-20-2019)

SDS-PAGE: 10 ug/ul RIPA lysate of whole cells 4-12% Bis-tris gradient gel Transfer: Immobilon-FL transfer membranes (Millipore) O/N at 30V, 4C Blocking: SEA Block Blocking Buffer 1h Primary Ab: DDDDK pRb Cat# 20543-1-AP O/N incubation at 4C Secondary Ab: IRDye 800CW Goat anti-Rabbit 1 h incubation at room temperature Lines on WB: 1. BioRad Precision Plus Protein standards 2. Whole cell lysate of HEK293t cells (negative transfection) 3. Whole cell lysate of HEK293t cells transfected with FLAG-tag construct 27 kDa 3. Whole cell lysate of HEK293t cells transfected with FLAG-tag construct 40 kDa

  • Applications: Western Blot,
  • Primary Antibody Dilution: 1:1000
  • Cell Tissue Type: HEK293t
DYKDDDDK tag Antibody Western Blot, validation (1:1000 dilution) in HEK293t (Cat no:20543-1-AP)
FH

Luce (Verified Customer) (11-12-2018)

  • Applications: Western Blot,Immunoprecipitation,
  • Cell Tissue Type: 10T1/2 cells