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 L5 antibody is 10-15 fold more sensitive than Sigma's M2 mouse ANTI-FLAG M2 ...read more
Immunocytochemistry/ Immunofluorescence: DYKDDDDK Epitope Tag Antibody (L5) [NBP1-06712] - Aanalysis of HeLa cells transfected with a FLAG encoded construct (top) or untransfected (bottom). Image from verified customer ...read more
Western Blot: DYKDDDDK Epitope Tag Antibody (L5) [NBP1-06712] - Western Blot on DYKDDDDK tagged SFO-p41 protein.
Western Blot: DYKDDDDK Epitope Tag Antibody (L5) [NBP1-06712] - Analysis of DYKDDDDK tagged AIFm2 protein (A), DYKDDDDK tagged IREB2 protein (B), and DYKDDDDK tagged PGC1A (C) using DYKDDDDK epitope tag antibody (L5) ...read more
Western Blot: DYKDDDDK Epitope Tag Antibody (L5) [NBP1-06712] - Detection of DDK in SUSD3 overexpression lysate using anti-DYKDDDDK Epitope Tag antibody followed by goat anti-rat IgG HRP conjugated secondary antibody ...read more

Product Details

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

Order Details

View Available Conjugates

DYKDDDDK Epitope Tag Antibody (L5) Summary

Immunogen
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)
Reviewed Applications
Read 6 Reviews rated 4.7
using
NBP1-06712 in the following applications:

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

  • 6 publications
  • IHC
    5 publications
  • 1 publication
  • IP
    4 publications
  • WB
    10 publications

Reactivity Notes

Tag specific. Drosophila reactivity reported in scientific literature (PMID: 26573957). Guinea Pig reactivity reported in scientific literature (PMID: 28651121).

Packaging, Storage & Formulations

Storage
Store at 4C. Do not freeze.
Buffer
No buffer
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-DYKDDDDK tag
  • anti-flag L5
  • anti-FLAG Tag
  • 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

This rat monoclonal DYKDDDDK antibody is a useful tool for the localization and characterization of DDDDK tagged proteins. Testing shows that this monoclonal antibody has 2-8 times more sensitivity than the standard FLAG antibody(TM) epitope tag antibody (1).

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)(29)

We have publications tested in 5 confirmed species: Human, Mouse, Rat, Drosophila, Guniea Pig.

We have publications tested in 5 applications: ICC/IF, IHC, IHC-Fr, IP, WB.


Filter By Application
ICC/IF
(6)
IHC
(5)
IHC-Fr
(1)
IP
(4)
WB
(10)
All Applications
Filter By Species
Human
(5)
Mouse
(2)
Rat
(1)
Drosophila
(2)
Guniea Pig
(1)
All Species
Showing Publications 1 - 10 of 29. Show All 29 Publications.
Publications using NBP1-06712 Applications Species
Yamamoto H, Umeda D, Matsumoto S, Kikuchi A. LDL switches the LRP6 internalization route from flotillin-dependent to clathrin-dependent in hepatic cells J. Cell. Sci. 2017 Aug 18 [PMID: 28821575] (ICC/IF) ICC/IF
Coleman S, Choi KY, McGregor A. Cytomegalovirus UL128 homolog mutants that form a pentameric complex produce virus with impaired epithelial and trophoblast cell tropism and altered pathogenicity in the guinea pig. Virology. 2017 Jun 23 [PMID: 28651121] (Guniea Pig) Guniea Pig
Hattori D, Aso Y, Swartz KJ et al. Representations of Novelty and Familiarity in a Mushroom Body Compartment. Cell May 18 2017 [PMID: 28502772]
Wreden CC, Meng JL, Feng W et al. Temporal Cohorts of Lineage-Related Neurons Perform Analogous Functions in Distinct Sensorimotor Circuits. Curr. Biol. May 22 2017 [PMID: 28502656]
Earley LF, Powers JM, Adachi K et al. Adeno-Associated Virus Assembly-Activating Protein Is Not an Essential Requirement for Capsid Assembly of AAV Serotypes 4, 5 and 11. J. Virol. Nov 16 2016 [PMID: 27852862]
Shirangi TR, Wong AM, Truman JW, Stern DL. Doublesex Regulates the Connectivity of a Neural Circuit Controlling Drosophila Male Courtship Song. Dev. Cell Jun 20 2016 [PMID: 27326931] (IHC) IHC
Yokoi N, Fukata Y, Sekiya A et al. Identification of PSD-95 Depalmitoylating Enzymes. J. Neurosci. Jun 15 2016 [PMID: 27307232]
Zhou B, Ritt DA, Morrison DK et al. Ck2alpha Maintains ERk Activity in a kinase-independent Manner to Promote Resistance to Inhibitors of RAF and MEk but not ERk in BRAF-mutant Melanoma. J. Biol. Chem. May 17 2016 [PMID: 27226552] (WB) WB
Li N, Subrahmanyan L, Smith E et al. Mutations in the Histone Modifier PRDM6 Are Associated with Isolated Nonsyndromic Patent Ductus Arteriosus. Am. J. Hum. Genet. Jun 2 2016 [PMID: 27181681] (WB, ICC/IF, Human) WB, ICC/IF Human
Lacin H, Truman JW. Lineage mapping identifies molecular and architectural similarities between the larval and adult Drosophila central nervous system. Elife. 2016 Mar 15 [PMID: 26975248] (IHC) IHC
Show All 29 Publications.

Reviews for DYKDDDDK Epitope Tag Antibody (NBP1-06712) (6) 4.76

Average Rating: 4.7
(Based on 6 reviews)
We have 6 reviews tested in 3 species: Human, Rat, Other.
We have 6 reviews tested in 3 application: IP, IF, WB.

Reviews using NBP1-06712:
Filter by Applications
IP
(1)
IF
(2)
WB
(3)
All Applications
Filter by Species
Human
(3)
Rat
(1)
Other
(2)
All Species
Images Ratings Applications Species Date Details
Immunoprecipitation DYKDDDDK Epitope Tag NBP1-06712
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reviewed by:
Anonymous
IP Human 03/30/2017
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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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reviewed by:
Anonymous
IF Rat 06/02/2015
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Summary

ApplicationImmunofluorescence
SpeciesRat
FileView PDF
Western Blot DYKDDDDK Epitope Tag NBP1-06712
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reviewed by:
Anonymous
WB Human 03/13/2015
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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
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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.
 
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.
 
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 - 8 of 8 FAQs).

  1. Are the FLAG Tag antibodies validated in Simple Western?
    • Yes, we offer a 2 FLAG Tag antibodies that have been tested in Simple Western: NB600-345 and NBP1-71705.
  2. Does FLAG Tag antibodies comes in lyophilized form?
    • The following FLAG Tag antibodies come in lyophilized form: NBP1-97401, NBP1-97399, NBP1-97400, NBP1-97402.
  3. What the theoretical molecular weight for FLAG Tag antibodies?
    • The TMW of FLAG Tag antibodies is approximately 150 kDa.
  4. 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.
  5. Will FLAG Tag antibodies recognizes FLAG at the C-terminal end?
    • Yes, FLAG Tag antibodies can recognize FLAG-tag at both the N- or C-terminal.
  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.
  8. What is the immunogen sequence of this FLAG Tag antibody?
    • A sequence within extracellular domain of Mouse Langerin. Binds to same epitope as Sigma's Anti-FLAG® M2 Antibody. The exact sequence is proprietary.

Secondary Antibodies

 

Isotype Controls

Other Available Formats

Biotin Labeled NBP1-06712B
DyLight 680 Labeled NBP1-06712FR
DyLight 755 Labeled NBP1-06712IR

Additional DYKDDDDK Epitope Tag Products

DYKDDDDK Epitope Tag NBP1-06712

Research Areas for DYKDDDDK Epitope Tag Antibody (NBP1-06712)

Find related products by research area.

Blogs on DYKDDDDK Epitope Tag.

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
4
4
2
3
0
2
0
1
0

Anonymous
03/30/2017
Application: IP
Species: Human

Anonymous
06/02/2015
Application: IF
Species: Rat

Anonymous
03/13/2015
Application: WB
Species: Human