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

Publications

292

CloneNo.

8H6A10

Reviews (3)

Cat no : 66008-4-Ig

Synonyms

Flag tag, Flag, DYKDDDDK, DDDDK tag, DDDDK

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• Validation Data Gallery
• Proteintech vs. Other Vendors

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  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.

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  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

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  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.

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  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.

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  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).

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  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 .

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Tested Applications

• Positive WB detected in: Transfected HEK-293 cells
• Positive IP detected in: Transfected HEK-293 cells
• Positive IF/ICC detected in: Transfected HEK-293 cells
• Positive FC (Intra) detected in: Transfected Flag-TDP43 HEK-293 cells

Recommended dilution

• 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)/ICC: IF/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.

Published Applications

• WB: See 207 publications below
• IHC: See 2 publications below
• IF: See 36 publications below
• IP: See 66 publications below
• CoIP: See 39 publications below
• ChIP: See 5 publications below
• RIP: See 2 publications below

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 Reactivity: human, mouse, rat, pig, canine, monkey, chicken, zebrafish, hamster, duck
• Host / Isotype: Mouse / IgG2b
• Class: Monoclonal
• Type: Antibody
• Immunogen: Peptide
• Full Name: DYKDDDDK tag
• RRID: AB_2918475
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC 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.

Publications

Species	Application	Title
	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
human	WB	Adv Sci (Weinh); Inhibition of CK2/ING4 Pathway Facilitates Non-Small Cell Lung Cancer Immunotherapy; Authors - Qian Gou
human	WB,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.

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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