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DYKDDDDK Epitope Tag Antibody (L5)

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Western Blot: DYKDDDDK Epitope Tag Antibody (L5) [NBP1-06712] - Analysis of DYKDDDDK tagged protein demonstrating that the rat monoclonal DYKDDDDK Epitope Tag Antibody (L5) [NBP1-06712] is 10-15 fold more sensitive than ...read more
Western Blot: DYKDDDDK Epitope Tag Antibody (L5) [NBP1-06712] - Analysis of DYKDDDDK tagged SFO-p41 protein with DYKDDDDK Epitope Tag Antibody (L5) [NBP1-06712]. Observed molecular weight ~46.
Western Blot: DYKDDDDK Epitope Tag Antibody (L5) [NBP1-06712] - Analysis of DYKDDDDK tagged AIFm2 protein (A), DYKDDDDK tagged IREB2 protein (B), and vector control (C) using DYKDDDDK epitope tag antibody (L5) ...read more
Immunocytochemistry/ Immunofluorescence: DYKDDDDK Epitope Tag Antibody (L5) [NBP1-06712] - Analysis of HeLa cells transfected with a FLAG encoded construct (top) or untransfected (bottom), using DYKDDDDK Epitope Tag ...read more
Immunohistochemistry: DYKDDDDK Epitope Tag Antibody (L5) [NBP1-06712] - Third instar larva of the genotype [hs-Flp; nrv2-Gal4/MCFO-2]. flp expression of the multicolor FlpOut construct was induced by 1 h 37C heat shock ...read more
Western Blot: DYKDDDDK Epitope Tag Antibody (L5) [NBP1-06712] - Detection of DDK in SUSD3 overexpression lysate using [NBP1-06712] followed by goat anti-rat IgG HRP conjugated secondary antibody (cat.# NB7115). Image ...read more
Immunocytochemistry/ Immunofluorescence: Rat Monoclonal DYKDDDDK Epitope Tag Antibody (L5) [NBP1-06712] - Analysis using DyLight 488 conjugated antibody (NBP1-06712G). Normal mouse prostate cells overexpressed with ...read more
SHP inhibits AhR transactivation of Pemt in response to feeding or FGF19 treatment. a, b Hepa1c1c7 cells were grown in low-glucose and serum-free media for 12 h, and transferred for 15 min to complete medium or ...read more

Product Details

Summary
Reactivity ETSpecies Glossary
Applications WB, ICC/IF, IHC, IP
Clone
L5
Clonality
Monoclonal
Host
Rat
Conjugate
Unconjugated

Order Details

DYKDDDDK Epitope Tag Antibody (L5) Summary

Immunogen
DYKDDDDK Epitope Tag Antibody (L5) was made to N-terminal DYKDDDDK-tagged extracellular domain of mouse Langerin. Binds to same epitope as Sigma's Anti-FLAG® M2 Antibody.
Epitope
DDDDK
Specificity
Binds to same epitope as Sigma's Anti-FLAGM2 Antibody. FLAG is a registered trademark of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co.
Isotype
IgG2a
Clonality
Monoclonal
Host
Rat
Purity
Tissue culture supernatant
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.

Applications/Dilutions

Dilutions
  • Immunocytochemistry/ Immunofluorescence 1:10-1:100. Use reported in multiple pieces of scientific literature
  • Immunohistochemistry 1:50-1:100. Use reported in multiple pieces of scientific literature
  • Immunohistochemistry-Frozen reported in scientific literature (PMID 24454782)
  • Immunoprecipitation reported in multiple pieces of scientific literature
  • Western Blot 1:500-1:1000. Use reported in multiple pieces of scientific literature
Theoretical MW
1.01 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Reviewed Applications
Read 10 Reviews rated 4.7
using
NBP1-06712 in the following applications:

Publications
Read Publications using
NBP1-06712 in the following applications:

  • EM
    1 publication
  • IB
    1 publication
  • 22 publications
  • 29 publications
  • IHC
    8 publications
  • 3 publications
  • IP
    6 publications
  • WB
    26 publications

Packaging, Storage & Formulations

Storage
Store at 4C. Do not freeze.
Buffer
Tissue culture supernatant
Preservative
0.1% Sodium Azide
Purity
Tissue culture supernatant

Alternate Names for DYKDDDDK Epitope Tag Antibody (L5)

  • anti-flag L5
  • DDDDK epitope tag
  • DDK
  • DYKDDDDK Epitope Tag
  • DYKDDDDK tag
  • ECS epitope tag
  • ECS tag
  • Enterokinase Cleavage Site epitope tag
  • Enterokinase Cleavage Site tag
  • epitope tag flag
  • FLAG (FLAG is a trademark of Sigma Aldrich)
  • FLAG Tag
  • Ty 1 Tag
  • xxxDDDDK epitope tag

Background

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.

Limitations

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.

Publications for DYKDDDDK Epitope Tag Antibody (NBP1-06712)(143)

We have publications tested in 3 confirmed species: Mouse, Drosophila, Epitope Tag.

We have publications tested in 8 applications: EM, IB, ICC/IF, IF/IHC, IHC, IHC-Fr, IP, WB.


Filter By Application
EM
(1)
IB
(1)
ICC/IF
(22)
IF/IHC
(29)
IHC
(8)
IHC-Fr
(3)
IP
(6)
WB
(26)
All Applications
Filter By Species
Mouse
(1)
Drosophila
(1)
Epitope Tag
(2)
All Species
Showing Publications 1 - 10 of 143. Show All 143 Publications.
Publications using NBP1-06712 Applications Species
Radwanska K, Pagano R, Salamian A et al. Arc controls alcohol cue relapse by a central amygdala mechanism Research Square 2022-08-22 [PMID: 36357670] (IHC-Fr) IHC-Fr
Baker CA, McKellar C, Pang R et al. Neural network organization for courtship-song feature detection in Drosophila Current biology : CB 2022-06-28 [PMID: 35793679]
Fujiwara T, Brotas M, Chiappe ME Walking strides direct rapid and flexible recruitment of visual circuits for course control in Drosophila Neuron 2022-05-05 [PMID: 35525243]
Sun L, Jiang RH, Ye WJ Et al. Recurrent circadian circuitry regulates central brain activity to maintain sleep Neuron 2022-05-07 [PMID: 35525241]

Details:
Citation using the Biotin version of this antibody.
Bayley R, Borel V, Moss RJ et al. H3K4 methylation by SETD1A/BOD1L facilitates RIF1-dependent NHEJ Molecular cell 2022-04-07 [PMID: 35439434]
Certel SJ, McCabe BD, Stowers RS. Et al. A conditional GABAergic synaptic vesicle marker for Drosophila J Neurosci Methods 2022-02-27 [PMID: 35219770] (IF/IHC)

Details:
Citation using the Janelia Fluor 549 version of this antibody.
IF/IHC
Bajar BT, Phi NT, Isaacman-Beck J et al. A discrete neuronal population coordinates brain-wide developmental activity Nature 2022-02-01 [PMID: 35140397] (IF/IHC) IF/IHC
Marshall ZD, Heckscher ES The role of Even-skipped in Drosophila larval somatosensory circuit assembly eNeuro 2022-01-10 [PMID: 35031555]
Huang S, Yamaji N, Sakurai G et al. A pericycle-localized silicon transporter for efficient xylem loading in rice The New phytologist 2022-01-12 [PMID: 35020209]
Chen C, Agrawal S, Mark B Et al. Functional architecture of neural circuits for leg proprioception in Drosophila Current biology : CB 2021-10-08 [PMID: 34637749] (IF/IHC) IF/IHC
Show All 143 Publications.

Reviews for DYKDDDDK Epitope Tag Antibody (NBP1-06712) (10) 4.710

Average Rating: 4.7
(Based on 10 reviews)
We have 10 reviews tested in 3 species: Human, Rat, Other.

Reviews using NBP1-06712:
Filter by Applications
WB
(7)
IP
(1)
IF
(2)
All Applications
Filter by Species
Human
(7)
Rat
(1)
Other
(2)
All Species
Images Ratings Applications Species Date Details
  5
reviewed by:
Verified Customer
WB Human 01/28/2022
View

Summary

ApplicationWestern Blot
Sample TestedHEK 293 cells
SpeciesHuman
LotL5
Western Blot DYKDDDDK Epitope Tag NBP1-06712
Enlarge
5
reviewed by:
Verified Customer
WB Human 04/04/2020
View

Summary

ApplicationWestern Blot
Sample Tested293T
SpeciesHuman
LotB-7
Western Blot DYKDDDDK Epitope Tag NBP1-06712
Enlarge
5
reviewed by:
Verified Customer
WB Human 12/27/2019
View

Summary

ApplicationWestern Blot
Sample Tested293 cell extract
SpeciesHuman
LotB-7
Western Blot DYKDDDDK Epitope Tag NBP1-06712
Enlarge
4
reviewed by:
Verified Customer
WB Human 02/18/2019
View

Summary

ApplicationWestern Blot
Sample Tested293T
SpeciesHuman
LotB-7
Immunoprecipitation DYKDDDDK Epitope Tag NBP1-06712
Enlarge
5
reviewed by:
Verified Customer
IP Human 03/30/2017
View

Summary

ApplicationImmunoprecipitation
Sample TestedLane 1: Flag-tagged protein tranfected HEK293 cell lysate pulled down by NBP1-06712,Lane 2: Untransfected HEK293 cells pulled down by NBP1-06712 (Control)
SpeciesHuman
LotB-1
Immunofluorescence DYKDDDDK Epitope Tag NBP1-06712
Enlarge
5
reviewed by:
Verified Customer
IF Rat 06/02/2015
View

Summary

ApplicationImmunofluorescence
SpeciesRat
Western Blot DYKDDDDK Epitope Tag NBP1-06712
Enlarge
5
reviewed by:
Verified Customer
WB Human 03/13/2015
View

Summary

ApplicationWestern Blot
Sample TestedSUSD3 Overexpression Lysate, NBL1-16623
SpeciesHuman
LotA-6
Western Blot DYKDDDDK Epitope Tag NBP1-06712
Enlarge
4
reviewed by:
Matt Stratton
WB Other 06/11/2014
View

Summary

ApplicationWestern Blot
Sample TestedCos 7 cells infected with adenovirus
SpeciesOther
  5
reviewed by:
Verified Customer
IF Human 05/27/2010
View

Summary

ApplicationImmunofluorescence
Sample TestedHek 293T
SpeciesHuman
LotA
  4
reviewed by:
Verified Customer
WB Other 03/31/2010
View

Summary

ApplicationWestern Blot
Sample TestedNovus TRUSS lysate (NBL1-17333)
SpeciesOther
LotA

Product General Protocols

View specific protocols for DYKDDDDK Epitope Tag Antibody (NBP1-06712): Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

Video Protocols

WB Video Protocol
ICC/IF Video Protocol

FAQs for DYKDDDDK Epitope Tag Antibody (NBP1-06712). (Showing 1 - 7 of 7 FAQs).

  1. Are the FLAG Tag antibodies validated in Simple Western?
    • Yes, we offer 3 FLAG Tag antibodies that have been tested in Simple Western: NBP1-71705, NBP1-97410, and MAB8529.
  2. What the theoretical molecular weight for FLAG Tag antibodies?
    • The TMW for DYKDDDDK epitope FLAG Tag is approximately 1.01 kDa.
  3. Will FLAG Tag antibodies detect internal FLAG tags? My FLAG sequence is sandwiched between two other protein sequences.
    • 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.
  4. Will FLAG Tag antibodies recognize FLAG at the C-terminal end?
    • Yes, FLAG Tag antibodies can recognize FLAG-tag at both the N- or C-terminal.
  5. Do FLAG Tag antibodies comes in lyophilized form?
    • The following FLAG Tag antibodies come in lyophilized form: NBP1-97393, NBP1-97401, NBP1-97399, NBP1-97400, and NBP1-97402.
  6. Upon the customer's inquiry, may we ask about the concentration of NBP1-06712?
    • NBP1-06712 is unpurified (tissue culture supernatant) and does not have a concentration.
  7. I ordered your NBP1-06712SS Rat Monoclonal Flag tag antibody. I have 2 questions: (1) Will this antibody recognize FLAG at the C-terminal end ? (2) Does my secondary need to be rat ? Or can it be mouse ?
    • This antibody will recognize FLAG-tag at N- or C-termnus. This antibody is a rat monoclonal antibody, so you will need to use an anti-rat secondary.

Secondary Antibodies

 

Isotype Controls

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By Jamshed Arslan Pharm.D. Glioblastoma is an aggressive type of cancer that forms from the star-shaped glial cells of the central nervous system, called astrocytes. Intriguingly, several genes linked to glioblasto...  Read full blog post.

Hoist the DYKDDDDK Epitope Tag
The DYKDDDDK Epitope Tag antibody was raised against a short, artificial sequence that is widely and commonly used as a protein fusion tag. It is pervasively present in a vast amount of scientific studies, and is a very powerful investigative research...  Read full blog post.

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Recent Reviews

4.7
5
7
4
3
3
0
2
0
1
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Verified Customer
01/28/2022
Application: WB
Species: Human

Verified Customer
04/04/2020
Application: WB
Species: Human

Verified Customer
12/27/2019
Application: WB
Species: Human