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

DYKDDDDK tag Monoclonal Antibody for WB, IF/ICC, FC (Intra), IP, ELISA

Host / Isotype

Mouse / IgG2b

Reactivity

recombinant protein and More (10)

Applications

WB, IHC, IF/ICC, FC (Intra), IP, CoIP, ChIP, RIP, ELISA

Conjugate

Unconjugated

CloneNo.

8H6A10

Cat no : 66008-4-Ig

Synonyms

Flag tag, Flag, DYKDDDDK, DDDDK tag, DDDDK


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  • Western Blot (WB) analysis of various lysates using DYKDDDDK tag Monoclonal antibody (Binds to FLAG® t (66008-4-Ig)

    WB analysis using 66008-4-Ig

    HEK-293 and different loading amount of DYKDDDDK tagged TDP-43 transfected HEK-293 cells were subjected to SDS PAGE followed by western blot with 66008-4-Ig (DYKDDDDK tag antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours. The membrane was re-blotted with GAPDH antibody (HRP-60004) as loading control.

  • Western Blot (WB) analysis of various lysates using DYKDDDDK tag Monoclonal antibody (Binds to FLAG® t (66008-4-Ig)

    WB analysis using 66008-4-Ig

    HEK-293 and DYKDDDDK-TDP43 transfected HEK-293 cells were subjected to SDS PAGE followed by western blot with 66008-4-Ig (DYKDDDDK tag antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.The membrane was re-blotted with GAPDH antibody (HRP-60004) as loading

  • Western Blot (WB) analysis of various lysates using DYKDDDDK tag Monoclonal antibody (Binds to FLAG® t (66008-4-Ig)

    WB analysis using 66008-4-Ig

    HEK-293 and DYKDDDDK-PCSK9 transfected HEK-293 cells were subjected to SDS PAGE followed by western blot with 66008-4-Ig (DYKDDDDKtag antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.The membrane was re-blotted with GAPDH antibody (HRP-60004) as loading control.

  • Immunoprecipitation (IP) experiment of Transfected HEK-293 cells using DYKDDDDK tag Monoclonal antibody (Binds to FLAG® t (66008-4-Ig)

    IP experiment of Transfected HEK-293 using 66008-4-Ig

    IP result of anti-DYKDDDDK tag (IP:66008-4-Ig, 5 ug; Detection:80010-1-RR 1:10000) with Transfected HEK-293T cells lysate 250 ug.

  • Immunofluorescence (IF) / fluorescent staining of Transfected HEK-293 cells using DYKDDDDK tag Monoclonal antibody (Binds to FLAG® t (66008-4-Ig)

    IF Staining of Transfected HEK-293 using 66008-4-Ig

    Immunofluorescent analysis of (-20°C Ethanol) fixed DYKDDDDK-TDP43 Transfected HEK-293 cells using DYKDDDDK tag antibody (66008-4-Ig, Clone: 8H6A10 ) at dilution of 1:2000 and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L).

  • Flow cytometry (FC) experiment of HEK-293 cells using DYKDDDDK tag Monoclonal antibody (Binds to FLAG® t (66008-4-Ig)

    FC experiment of HEK-293 using 66008-4-Ig

    1x10^6 HEK-293 cells (dash lines) and DYKDDDDK-TDP43 Transfected HEK-293 cells (solid lines) were intracellularly stained with 0.2 μg DYKDDDDK tag Monoclonal antibody (66008-4-Ig, Clone: 8H6A10, red) and CoraLite488-conjugated Goat Anti-Mouse IgG(H+L) (Cat.NO. SA00013-1). Mouse IgG2b isotype control (66360-3-Ig, Clone: 11B8C4, blue) was parallel stained as control. Cells were fixed with 4% PFA .

"DYKDDDDK tag Antibodies" Comparison

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Tested Applications

Positive WB detected inTransfected HEK-293 cells
Positive IP detected inTransfected HEK-293 cells
Positive IF/ICC detected inTransfected HEK-293 cells
Positive FC (Intra) detected inTransfected Flag-TDP43 HEK-293 cells

Recommended dilution

ApplicationDilution
Western Blot (WB)WB : 1:5000-1:50000
Immunoprecipitation (IP)IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
Immunofluorescence (IF)/ICCIF/ICC : 1:1000-1:4000
Flow Cytometry (FC) (INTRA)FC (INTRA) : 0.20 ug per 10^6 cells in a 100 µl suspension
It is recommended that this reagent should be titrated in each testing system to obtain optimal results.
Sample-dependent, Check data in validation data gallery.

Product Information

66008-4-Ig targets DYKDDDDK tag in WB, IHC, IF/ICC, FC (Intra), IP, CoIP, ChIP, RIP, ELISA applications and shows reactivity with recombinant protein samples.

Tested Reactivity recombinant protein
Cited Reactivityhuman, mouse, pig, canine, monkey, chicken, zebrafish, bovine, hamster, duck
Host / Isotype Mouse / IgG2b
Class Monoclonal
Type Antibody
Immunogen Peptide
Full Name DYKDDDDK tag
Gene Symbol
Gene ID (NCBI)
RRIDAB_2918475
Conjugate Unconjugated
Form Liquid
Purification MethodProtein A purification
Storage Buffer PBS with 0.02% 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. 20ul sizes contain 0.1% BSA.

Background Information

DYKDDDDK Tag ( Equivalent To FLAG Antibody From Sigma ) with the following sequence DYKDDDDK, is a hydrophilic tag for recombinant protein technology. Tags can be used as a tool to localize gene products in a variety of cell types, study proteins topology, and also help to identify and characterize new, low abundance or poorly immunogenic proteins. Due to its high hydrophilic character, the DKYDDDDK tag is likely to be located on the surface of a fusion protein, which enables the tag to be accessible for antibodies. DYKDDDDK Tag Antibody is generated against 1xDYKDDDDK tag (DYKDDDDK) and can recognize protein containing one or more DYKDDDDK tags, independently on N-terminal, C-terminal or internal regions of the target protein. Anti-FLAG is a registered trademark of Sigma-Aldrich Biotechnology.

Protocols

Product Specific Protocols
WB protocol for DYKDDDDK tag antibody 66008-4-IgDownload protocol
IF protocol for DYKDDDDK tag antibody 66008-4-IgDownload protocol
IP protocol for DYKDDDDK tag antibody 66008-4-IgDownload protocol
Standard Protocols
Click here to view our Standard Protocols

Publications

SpeciesApplicationTitle
ChIP

Blood

Functional, structural and molecular characterizations of leukemogenic driver MEF2D-HNRNPUL1 fusion.

Authors - Ming Zhang
WB

Nat Commun

UV-B irradiation-activated E3 ligase GmILPA1 modulates gibberellin catabolism to increase plant height in soybean

Authors - Jiaqi Sun
IP

Nat Commun

Homodimer-mediated phosphorylation of C/EBPα-p42 S16 modulates acute myeloid leukaemia differentiation through liquid-liquid phase separation

Authors - Dongmei Wang
humanWB

Adv Sci (Weinh)

Inhibition of CK2/ING4 Pathway Facilitates Non-Small Cell Lung Cancer Immunotherapy

Authors - Qian Gou
humanWB,IP

Cell Death Differ

Lysine methylation of PPP1CA by the methyltransferase SUV39H2 disrupts TFEB-dependent autophagy and promotes intervertebral disc degeneration

Authors - Huaizhen Liang
WB,IF,IP

Cell Death Differ

Deubiquitinase OTUD6A in macrophages promotes intestinal inflammation and colitis via deubiquitination of NLRP3

Authors - Xin Liu

Reviews

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


FH

Boyan (Verified Customer) (12-12-2022)

worked very well

  • Applications: Western Blot, Immunofluorescence
FH

Chun (Verified Customer) (06-19-2022)

This antibody

  • Applications: Western Blot
  • Primary Antibody Dilution: 1:1,000
FH

Céline (Verified Customer) (03-29-2022)

I have compared with anti-Flag M2 from Sigma in an IP experiment and I found the M2 antibody better.

  • Applications: Immunoprecipitation
  • Primary Antibody Dilution: 1/100
  • Cell Tissue Type: Hela cells