DYKDDDDK Epitope Tag Antibody (L5)

Images

 
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
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

Product Details

Summary
Reactivity All-NASpecies Glossary
Applications WB, ICC/IF, IHC, IHC-Fr, IP
Clone
L5
Clonality
Monoclonal
Host
Rat
Conjugate
Unconjugated

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
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
  • Western Blot 1:500-1:1000
  • Immunocytochemistry/Immunofluorescence 1:10-1:100
  • Immunohistochemistry 1:50-1:100
  • Immunohistochemistry-Frozen
  • Immunoprecipitation 1:10-1:500
Application Notes
This DYKDDDDK Epitope Tag antibody is useful for Immunocytochemistry/Immunofluorescence, Immunoprecipitation, Western blot and Immunohistochemistry. Use in Immunohistochemistry-Frozen reported in scientific literature (PMID 24454782). Use in ICC/IF, Immunohistochemistry, Immunoprecipitation, and Western Blot 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 9 Reviews rated 4.7
using
NBP1-06712 in the following applications:

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

  • IB
    1 publication
  • 12 publications
  • IHC
    25 publications
  • 1 publication
  • IP
    5 publications
  • WB
    18 publications

Reactivity Notes

Tag specific. Reactivity reported in multiple pieces of scientific literature. (PMID: 25896325), (PMID: 28651121).

Packaging, Storage & Formulations

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

Notes

Binds to same epitope as Sigma's Anti-FLAGM2 Antibody. FLAG is a registered trademark of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co.

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)(81)

We have publications tested in 3 confirmed species: Drosophila, Insect, Mammal.

We have publications tested in 6 applications: IB, ICC/IF, IHC, IHC-Fr, IP, WB.


Filter By Application
IB
(1)
ICC/IF
(12)
IHC
(25)
IHC-Fr
(1)
IP
(5)
WB
(18)
All Applications
Filter By Species
Drosophila
(2)
Insect
(2)
Mammal
(1)
All Species
Showing Publications 1 - 10 of 81. Show All 81 Publications.
Publications using NBP1-06712 Applications Species
Oh Y, Lai JS, Min S et al. Periphery signals generated by Piezo-mediated stomach stretch and Neuromedin-mediated glucose load regulate the Drosophila brain nutrient sensor Neuron May 13 2021 [PMID: 34015253]
Hiramoto A, Jonaitis J, Niki S et al. Regulation of coordinated muscular relaxation in Drosophila larvae by a pattern-regulating intersegmental circuit Nature communications May 19 2021 [PMID: 34011945] (IHC) IHC
Ding G, Li X, Hou X et al. REV-ERB in GABAergic neurons controls diurnal hepatic insulin sensitivity Nature Mar 24 2021 [PMID: 33762728]
Kitano M, Kizuka Y, Sobajima T, et al. Rab11-mediated post-Golgi transport of the sialyltransferase ST3GAL4 suggests a new mechanism for regulating glycosylation The Journal of biological chemistry Jan 29 2021 [PMID: 33524390]
Yano T, Tsukita K, Kanoh H et al. A microtubule-LUZP1 association around tight junction promotes epithelial cell apical constriction The EMBO journal Dec 21 2020 [PMID: 33346378] (WB) WB
Morimoto MM, Nern A, Zhao A et al. Spatial readout of visual looming in the central brain of Drosophila eLife Nov 18 2020 [PMID: 33205753]
Antonioli M, Pagni B, Vescovo T et al. HPV sensitizes OPSCC cells to cisplatin-induced apoptosis by inhibiting autophagy through E7-mediated degradation of AMBRA1 Autophagy Nov 23 2020 [PMID: 33172332] (IP) IP
Hampel S, Eichler K, Yamada D et al. Distinct subpopulations of mechanosensory chordotonal organ neurons elicit grooming of the fruit fly antennae Elife Oct 26 2020 [PMID: 33103999] (ICC/IF)

Details:
Immunofluorescence staining of Drosophila brains
ICC/IF
Kottmeier R, Bittern J, Schoofs A et al. Wrapping glia regulates neuronal signaling speed and precision in the peripheral nervous system of Drosophila Nature communications Sep 8 2020 [PMID: 32901033] (IHC) IHC
Sampson MM, Myers Gschweng KM, Hardcastle BJ et al. Serotonergic modulation of visual neurons in Drosophila melanogaster. PLoS Genet. Aug 31 2020 [PMID: 32866139] (IHC) IHC
Show All 81 Publications.

Reviews for DYKDDDDK Epitope Tag Antibody (NBP1-06712) (9) 4.79

Average Rating: 4.7
(Based on 9 reviews)
We have 9 reviews tested in 3 species: Human, Rat, Other.

Reviews using NBP1-06712:
Filter by Applications
WB
(6)
IP
(1)
IF
(2)
All Applications
Filter by Species
Human
(6)
Rat
(1)
Other
(2)
All Species
Images Ratings Applications Species Date Details
Western Blot DYKDDDDK Epitope Tag NBP1-06712
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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
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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
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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
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5
reviewed by:
Verified Customer
IF Rat 06/02/2015
View

Summary

ApplicationImmunofluorescence
SpeciesRat
FileView PDF
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

Blocking

Blocking Details2% cold fish geletin in TBS buffer, room temperature for 1hour

Primary Anitbody

Dilution Ratio1:1000 overnight at 4C in blocking buffer

Secondary Antibody

Secondary Descriptionanti-rat HRP Novus NB7115
Secondary Manufacturer Cat#NB7115
Secondary Concentration1:5000

Details

Detection Notes15-second exposure shows strong band at correct MW.

Comments

CommentsThis DDK tag antibody works very well. The only thing needs attention is that it is made in rat so needs a anti-rat secondary antibody.
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
CommentsHas also worked for immunoprecipitation in our hands.

Blocking

Blocking Details5% milk in TBST at room temp for 1 hour

Primary Anitbody

Dilution Ratio1:1000 dilution, overnight incubation, 4C, 2.5% BSA/TBST

Secondary Antibody

Secondary Descriptiongoat anti Rat IgG-HRP
Secondary Manufacturer Cat#sc-2065
Secondary Concentration1:2000

Details

Detection NotesChemiluninescent substrate - West PICO

Comments

CommentsHas also worked for immunoprecipitation in our hands.
  5
reviewed by:
IF Human 05/27/2010
View

Summary

ApplicationImmunofluorescence
Sample TestedHek 293T
SpeciesHuman
LotA
CommentsHek 293T cells were transfected with an N-terminal triple FLAG tagged (containing only one epitope for the antibody) cytoplasmic portion of KCNE1, fixed with -20oC methanol, stained with 40x dilute anti-DYKDDDDK (L5), AlexaFluor goat antirat secondary, and observed at 60x on a fluorescence microscope. A high level of transfection with the plasmid appears to make the cell sick.

Primary Anitbody

Dilution RatioPrimary Ab Incubation Dilution Buffer: Tris Tween buffered saline (TTBS), Primary Ab Incubation Time: 1hr, Incubation Temp: RT

Secondary Antibody

Secondary DescriptionSecondary Ab Dilution: 1:1000

Details

Detection NotesDetection Method: Fluorescence@488nm, Control: untransfected (unstained) cells among transfected cells

Comments

CommentsHek 293T cells were transfected with an N-terminal triple FLAG tagged (containing only one epitope for the antibody) cytoplasmic portion of KCNE1, fixed with -20oC methanol, stained with 40x dilute anti-DYKDDDDK (L5), AlexaFluor goat antirat secondary, and observed at 60x on a fluorescence microscope. A high level of transfection with the plasmid appears to make the cell sick.
  4
reviewed by:
WB Other 03/31/2010
View

Summary

ApplicationWestern Blot
Sample TestedNovus TRUSS lysate (NBL1-17333)
SpeciesOther
LotA
CommentsI used this product with the Novus TRUSS lysate (NBL1-17333) and a band was detected at 90 kDa as expected. The negative control provided with the TRUSS lysate did not have a band at 90 kDa. There were minimal background bands.

Blocking

Blocking DetailsBlocking Buffer: 5% Blotto, Blocking Time: 1 hour, Blocking Temp: Room temperature

Primary Anitbody

Dilution RatioPrimary Ab Dilution Ratio: 1:1,000, Primary Ab Incubation Time: overnight, Primary Ab Incubation Temp: 4 degrees Celsius

Comments

CommentsI used this product with the Novus TRUSS lysate (NBL1-17333) and a band was detected at 90 kDa as expected. The negative control provided with the TRUSS lysate did not have a band at 90 kDa. There were minimal background bands.

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

Other Available Formats

Alexa Fluor 405 NBP1-06712AF405
Alexa Fluor 488 NBP1-06712AF488
Alexa Fluor 532 NBP1-06712AF532
Alexa Fluor 594 NBP1-06712AF594
Alexa Fluor 647 NBP1-06712AF647
Alexa Fluor 700 NBP1-06712AF700
Alexa Fluor 750 NBP1-06712AF750
Biotin NBP1-06712B
DyLight 350 NBP1-06712UV
DyLight 405 NBP1-06712V
DyLight 488 NBP1-06712G
DyLight 550 NBP1-06712R
DyLight 594 NBP1-06712DL594
DyLight 650 NBP1-06712C
DyLight 680 NBP1-06712FR
DyLight 755 NBP1-06712IR
FITC NBP1-06712F
HRP NBP1-06712H
Janelia Fluor 549 NBP1-06712JF549
Janelia Fluor 646 NBP1-06712JF646

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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
6
4
3
3
0
2
0
1
0

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

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

Verified Customer
02/18/2019
Application: WB
Species: Human