DYKDDDDK Epitope Tag Antibody (L5)

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 antibody.

Western Blot: DYKDDDDK Epitope Tag Antibody (L5) [NBP1-06712] - 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 vector control (C) using DYKDDDDK epitope tag antibody (L5) (NBP1-06712) at 2 ug/ml.

Immunocytochemistry/Immunofluorescence: DYKDDDDK Epitope Tag Antibody (L5) [NBP1-06712] - Analysis of HeLa cells transfected with a FLAG encoded construct (top) or untransfected (bottom). Image from verified customer review.

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 (cat.# NB7115). Image from verified customer review.

• Reactivity: Hu, Dr, GP, NA
• Applications: WB, ICC/IF, IHC, IHC-Fr, IP
• Clone: L5
• Clonality: Monoclonal
• Host: Rat
• Conjugate: Unconjugated

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

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)

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: 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-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)(48)

Publications using NBP1-06712

• Esteves P, Dard L, Brillac A et al. Nuclear control of lung cancer cells migration, invasion and bioenergetics by eukaryotic translation initiation factor 3F Oncogene [PMID: 31527668] (WB, Human)
• Napit PR, Ali MH, Shakya M et al. Neurogenetic dissection of the Drosophila lateral horn reveals major outputs, diverse behavioural functions, and interactions with the mushroom body Elife May 21 2019 [PMID: 31112130] (IHC, Drosophila)
• McKellar CE, Lillvis JL, Bath DE et al. Threshold-Based Ordering of Sequential Actions during Drosophila Courtship. Curr. Biol. Jan 7 2019 [PMID: 30661796] (IHC, Drosophila)
• Barnhart EL, Wang IE, Wei H et al. Sequential Nonlinear Filtering of Local Motion Cues by Global Motion Circuits. Neuron Sep 6 2018 [PMID: 30220510] (IHC)
• Chang YC, Tsao CK, Sun YH. Temporal and spatial order of photoreceptor and glia projections into optic lobe in Drosophila Sci Rep Aug 23 2018 [PMID: 30140062] (IHC, Drosophila)
• Gupta A, Ajith A, Singh S et al. PAK2-c-Myc-PKM2 axis plays an essential role in head and neck oncogenesis via regulating Warburg effect Cell Death Dis Aug 1 2018 [PMID: 30068946] (WB, Human)
• Dolan M, Frechter S, Bates A et al. Neurogenetic dissection of the Drosophila innate olfactory processing center. bioRxiv Sep 12 2018 (IHC, Insect)
• Losier TT, Akuma M, McKee-Muir OC et al. AMPK Promotes Xenophagy through Priming of Autophagic Kinases upon Detection of Bacterial Outer Membrane Vesicles Cell Rep Feb 19 2019 [PMID: 30784596] (WB)
• Andrade I, Riebli N, Nguyen B et al. Developmentally arrested precursors of pontine neurons establish an embryonic blueprint of the Drosophila central complex bioRxiv 2018 Apr 11 [PMID: 30661802] (IHC, Drosophila)
• Cosmanescu F, Katsamba PS, Sergeeva AP et al. Neuron-Subtype-Specific Expression, Interaction Affinities, and Specificity Determinants of DIP/Dpr Cell Recognition Proteins. Neuron. Nov 12 2018 [PMID: 30467080] (Drosophila)
• Wolff T, Rubin GM. Neuroarchitecture of the Drosophila central complex: A catalog of nodulus and asymmetrical body neurons and a revision of the protocerebral bridge catalog J. Comp. Neurol. Nov 1 2018 [PMID: 30084503] (IHC)
• Courgeon M, Desplan C Coordination between stochastic and deterministic specification in the Drosophila visual system bioRxiv Jul 11 1905 (IHC, Drosophila)
• Ribeiro IMA, Drews M, Bahl A et al. Visual Projection Neurons Mediating Directed Courtship in Drosophila. Cell. Jul 26 2018 [PMID: 30033367] (ICC/IF)
• Pan C, Jin L, Wang X et al. Characterization of Post-Translational Modification of ATG16L1 in Antibacterial Autophagy Thesis (WB, Mammal)
• Konstantinides N, Kapuralin K, Fadil C et al. Phenotypic Convergence: Distinct Transcription Factors Regulate Common Terminal Features Cell Jun 9 2018 [PMID: 29909983] (Drosophila)
• Imaimatsu K, Fujii W, Hiramatsu R et al. CRISPR/Cas9-mediated knock-in of the murine Y chromosomal Sry gene. J. Reprod. Dev. Apr 14 2018 [PMID: 29657232] (ICC/IF)
• Xie T, Ho MCW, Liu Q et al. A Genetic Toolkit for Dissecting Dopamine Circuit Function in Drosophila. Cell Rep Apr 10 2018 [PMID: 29642019] (ICC/IF)
• Saumweber T, Rohwedder A, Schleyer M et al. Functional architecture of reward learning in mushroom body extrinsic neurons of larval Drosophila. Nat Commun. Mar 16 2018 [PMID: 29549237] (Drosophila)
• Tocker AM, Durocher E, Jacob kD et al. The Scaffolding Protein IQGAP1 Interacts with NLRC3 and Inhibits Type I IFN Production. J. Immunol. Oct 15 2017 [PMID: 28864474] (ICC/IF, Human)
• 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)
• 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)
• 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)
• 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)
• 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)
• 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)
• Ichinose T, Aso Y, Yamagata N et al. Reward signal in a recurrent circuit drives appetitive long-term memory formation. Elife. 2015 Nov 17 [PMID: 26573957] (Drosophila)
• Hampel S, Franconville R, Simpson JH, Seeds AM. A neural command circuit for grooming movement control. Elife 2015 [PMID: 26344548] (IHC)
• Ohyama T, Schneider-Mizell CM, Fetter RD et al. A multilevel multimodal circuit enhances action selection in Drosophila Nature 2015 Apr 30 [PMID: 25896325] (IHC, Rat)
• Earley LF, Kawano Y, Adachi K et al. Identification and characterization of nuclear and nucleolar localization signals in the adeno-associated virus serotype 2 assembly-activating protein. J. Virol. 2014 Dec 31 [PMID: 25552709] (ICC/IF, Human)
• Rampal R, Alkalin A, Madzo J et al. DNA Hydroxymethylation Profiling Reveals that WT1 Mutations Result in Loss of TET2 Function in Acute Myeloid Leukemia. Cell Rep. 2014 Dec 11 [PMID: 25482556] (IP, WB)
• Wolff T, Iyer Na, Rubin Gm. Neuroarchitecture and neuroanatomy of the Drosophila central complex: A GAL4-based dissection of protocerebral bridge neurons and circuits. J. Comp. neurol. 2014 Nov 07 [PMID: 25380328] (IHC, Drosophila)
• Oliverio S, Corazzari M, Sestito C et al. The spermidine analogue GC7 (N1-guanyl-1,7-diamineoheptane) induces autophagy through a mechanism not involving the hypusination of eIF5A. Amino Acids. 2014 Sep 14 [PMID: 25218134]
• Jun LS, Showalter AD, Ali N et al. A Novel Humanized GLP-1 Receptor Model Enables Both Affinity Purification and Cre-LoxP Deletion of the Receptor. PLoS ONE 41732 [PMID: 24695667] (WB)
• Andreou M, Yan CY, Skourides PA. 40LoVe and Samba are involved in Xenopus neural development and functionally distinct from hnRNP AB. PLoS ONE 2014 Jan 23 [PMID: 24454782] (IHC-Fr, WB)
• Ohkawa T, Fukata Y, Yamasaki M et al. Autoantibodies to Epilepsy-Related LGI1 in Limbic Encephalitis Neutralize LGI1-ADAM22 Interaction and Reduce Synaptic AMPA Receptors. J Neurosci. 2013 Nov 13 [PMID: 24227725] (ICC/IF)
• Nam G, Lee YK, Lee HY et al. Interaction of CD99 with Its Paralog CD99L2 Positively Regulates CD99L2 Trafficking to Cell Surfaces. J Immunol. 2013 Dec 1 [PMID: 24133166] (IP)
• Gouttenoire EA, Lupo V, Calpena E et al. Sh3tc2 deficiency affects neuregulin-1/ErbB signaling. Glia 2013 Apr 2 [PMID: 23553667] (ICC/IF)
• Abdel-Wahab O, Adli M, Lafave LM et al. ASXL1 Mutations Promote Myeloid Transformation through Loss of PRC2-Mediated Gene Repression Cancer Cell 2012 Aug 14 [PMID: 22897849] (IP, Human)
• Park YK, Park H. Differentiated embryo chondrocyte 1 (DEC1) represses PPAR?2 gene through interacting with CCAAT/enhancer binding protein ? (C/EBP?). Mol Cells. 2012 May 18. [PMID: 22610404] (WB, Mouse)
• Kuver A, Shen H, Smith SS. Regulation of the surface expression of ?4?2? GABA(A) receptors by high efficacy states. Brain Res. 2012 May 2. [PMID: 22609410] (WB, Human)
• Hubbi ME, Luo W, Baek JH et al. MCM proteins are negative regulators of hypoxia-inducible factor 1 Mol Cell 2011 Jun 10 [PMID: 21658608] (WB, IP)
• Idoyaga, J et al. Cutting edge: langerin/CD207 receptor on dendritic cells mediates efficient antigen presentation on MHC I and II products in vivo. J Immunol;180(6):3647-50. 2008 Mar 15. [PMID: 18322168] (WB, Mouse)
• Andreou MS. A study of 40LoVe's role in xenopus development and the development of a novel intein based method for the in vivo conjugation of quantum dots to target proteins. Thesis. 2014 Jun 01 (ICC/IF)
• Park, SH et al. Generation and application of new rat monoclonal antibodies against synthetic FLAG and OLLAS tags for improved immunodetection. J Immunol Methods;331(1-2):27-38. 2008 Feb 29. [PMID: 18054954] (WB, Human)

Reviews for DYKDDDDK Epitope Tag Antibody (NBP1-06712) (7)

Average Rating: 4.6

(Based on 7 reviews)

We have 7 reviews tested in 3 species: Human, Rat, Other.

reviewed by: Anonymous WB Human 02/18/2019

Summary

• Application: Western Blot
• Sample Tested: 293T
• Species: Human
• Lot: B-7

reviewed by: Anonymous IP Human 03/30/2017

Summary

• Application: Immunoprecipitation
• Sample Tested: Lane 1: Flag-tagged protein tranfected HEK293 cell lysate pulled down by NBP1-06712,Lane 2: Untransfected HEK293 cells pulled down by NBP1-06712 (Control)
• Species: Human
• Lot: B-1

reviewed by: Anonymous IF Rat 06/02/2015

Summary

• Application: Immunofluorescence
• Species: Rat
• File: View PDF

reviewed by: Anonymous WB Human 03/13/2015

Summary

• Application: Western Blot
• Sample Tested: SUSD3 Overexpression Lysate, NBL1-16623
• Species: Human
• Lot: A-6

Blocking

• Blocking Details: 2% cold fish geletin in TBS buffer, room temperature for 1hour

Primary Anitbody

• Dilution Ratio: 1:1000 overnight at 4C in blocking buffer

Secondary Antibody

• Secondary Description: anti-rat HRP Novus NB7115
• Secondary Manufacturer Cat#: NB7115
• Secondary Concentration: 1:5000

Details

• Detection Notes: 15-second exposure shows strong band at correct MW.

Comments

• Comments: This 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.

reviewed by: Matt Stratton WB Other 06/11/2014

Summary

• Application: Western Blot
• Sample Tested: Cos 7 cells infected with adenovirus
• Species: Other
• Comments: Has also worked for immunoprecipitation in our hands.

Blocking

• Blocking Details: 5% milk in TBST at room temp for 1 hour

Primary Anitbody

• Dilution Ratio: 1:1000 dilution, overnight incubation, 4C, 2.5% BSA/TBST

Secondary Antibody

• Secondary Description: goat anti Rat IgG-HRP
• Secondary Manufacturer Cat#: sc-2065
• Secondary Concentration: 1:2000

Details

• Detection Notes: Chemiluninescent substrate - West PICO

Comments

• Comments: Has also worked for immunoprecipitation in our hands.

reviewed by: IF Human 05/27/2010

Summary

• Application: Immunofluorescence
• Sample Tested: Hek 293T
• Species: Human
• Lot: A
• Comments: Hek 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 Ratio: Primary Ab Incubation Dilution Buffer: Tris Tween buffered saline (TTBS), Primary Ab Incubation Time: 1hr, Incubation Temp: RT

Secondary Antibody

• Secondary Description: Secondary Ab Dilution: 1:1000

Details

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

Comments

• Comments: Hek 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

Summary

• Application: Western Blot
• Sample Tested: Novus TRUSS lysate (NBL1-17333)
• Species: Other
• Lot: A
• Comments: I 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 Details: Blocking Buffer: 5% Blotto, Blocking Time: 1 hour, Blocking Temp: Room temperature

Primary Anitbody

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

Comments

• Comments: I 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.

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.