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Tags & Cell Markers Epitope Tags DDDDK Tag

Anti-DDDDK tag (Binds to FLAG® tag sequence)抗体(ab1162)

  • Datasheet
  • SDS
Reviews (15)Q&A (6)References (114)

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Immunocytochemistry - Anti-DDDDK tag (Binds to FLAG® tag sequence) antibody (ab1162)
  • Immunocytochemistry - Anti-DDDDK tag (Binds to FLAG® tag sequence) antibody (ab1162)
  • Immunocytochemistry - Anti-DDDDK tag (Binds to FLAG® tag sequence) antibody (ab1162)

Key features and details

  • Rabbit polyclonal to DDDDK tag (Binds to FLAG® tag sequence)
  • Suitable for: ICC, WB, ELISA
  • Reacts with: Species independent
  • Isotype: IgG

选择批间可重复性更高的重组抗体

Product image
Anti-DDDDK tag (Binds to FLAG® tag sequence) antibody [EPR20018-251] (ab205606)
  • 研究可靠 —— 各批次间结果一致且可重复
  • 长期批量供应 —— 采用重组技术,可实现快速生产
  • 首次实验即可成功 —— 经过大量验证确认了特异性
  • 符合伦理标准 —— 产品不含动物成分

概述

  • 产品名称

    Anti-DDDDK tag (Binds to FLAG® tag sequence)抗体
    参阅全部 DDDDK tag (Binds to FLAG® tag sequence) 一抗
  • 描述

    兔多克隆抗体to DDDDK tag (Binds to FLAG® tag sequence)
  • 宿主

    Rabbit
  • 经测试应用

    适用于: ICC, WB, ELISAmore details
  • 种属反应性

    与反应: Species independent
  • 免疫原

    Synthetic peptide corresponding to DDDDK tag (Binds to FLAG® tag sequence). Peptide DYKDDDDK conjugated to KLH. This sequence represents the Enterokinase Cleavage Site.

  • 常规说明

    Epitope tags provide a method to localize gene products in a variety of cell types, study the topology of proteins and protein complexes, identify associated proteins, and characterize newly identified, low abundance or poorly immunogenic proteins when protein specific antibodies are not available. Tagging with xxxDDDDK may be done at the N-terminus, N-terminus preceded by a methionine residue, C-terminus, and in internal positions of the target protein. The small size of the epitope tag and its high hydrophilicity tend to decrease the possibility of interference with protein expression, proteolytic maturation, antigenicity and function. The enterokinase cleavage site allows it to be completely removed from the purified fusion proteins.

    FLAG® is a registered trade mark of Sigma Aldrich Biotechnology LP. It is used here for informational purposes only.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

性能

  • 形式

    Liquid
  • 存放说明

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • 存储溶液

    pH: 7.20
    Preservative: 0.1% Sodium azide
    Constituent: PBS
  • Concentration information loading...
  • 纯度

    Immunogen affinity purified
  • 克隆

    多克隆
  • 同种型

    IgG
  • 研究领域

    • Tags & Cell Markers
    • Epitope Tags
    • DDDDK Tag

相关产品

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab1162于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
ICC (1)
Use at an assay dependent concentration.
WB (9)
Use a concentration of 0.2 - 1 µg/ml.

Antibody was analyzed by western blot using various amino-terminal and carboxy-terminal DDDDK fusion proteins and Invitrogen Positope" R900-40. A 1/5000 dilution gave bands against 0.1, 1.0 and 10 ng of the fusion proteins and 100 ng and 25 ng of the Positope.

ELISA
Use a concentration of 1 - 5 µg/ml.

As coating antibody in sELISA.

说明
ICC
Use at an assay dependent concentration.
WB
Use a concentration of 0.2 - 1 µg/ml.

Antibody was analyzed by western blot using various amino-terminal and carboxy-terminal DDDDK fusion proteins and Invitrogen Positope" R900-40. A 1/5000 dilution gave bands against 0.1, 1.0 and 10 ng of the fusion proteins and 100 ng and 25 ng of the Positope.

ELISA
Use a concentration of 1 - 5 µg/ml.

As coating antibody in sELISA.

靶标

  • 相关性

    This is a useful tool for the localisation and characterisation of DDDDK tagged proteins (Binds to FLAG® tag sequence).
  • 别名

    • DDDDK epitope tag antibody
    • DDDK antibody
    • ddk antibody
    • DYKDDDDK (also known as the FLAG® tag sequence) antibody
    • DYKDDDDK antibody
    • DYKDDDDK epitope tag antibody
    • DYKDDDDK tag antibody
    • ECS epitope tag antibody
    • ECS tag antibody
    • Enterokinase Cleavage Site epitope tag antibody
    • Enterokinase Cleavage Site tag antibody
    • FLAG® antibody
    • FLAG® tag antibody
    see all

图片

  • Immunocytochemistry - Anti-DDDDK tag (Binds to FLAG® tag sequence) antibody (ab1162)
    Immunocytochemistry - Anti-DDDDK tag (Binds to FLAG® tag sequence) antibody (ab1162)

    Immunofluorescent analysis of A293 cells labeling FLAG®-VPS35 with ab1162 at 10 μg/ml. The nuclear counter stain is DAPI (blue).

  • Immunocytochemistry - Anti-DDDDK tag (Binds to FLAG® tag sequence) antibody (ab1162)
    Immunocytochemistry - Anti-DDDDK tag (Binds to FLAG® tag sequence) antibody (ab1162)This image is courtesy of an anonymous Abreview
    ab1162 staining DDDDK tag in hTERT-RPE1 by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with Paraformaldehyde, permeabilized with Saponin and blocked with 0.02% BSA for 20 minutes at 18°C. Samples were incubated with primary antibody (1/100) for 45 minutes at 18°C. A Cy3®-conjugated Donkey anti-rabbit polyclonal was used as the secondary antibody.

    See Abreview

  • Immunocytochemistry - Anti-DDDDK tag (Binds to FLAG® tag sequence) antibody (ab1162)
    Immunocytochemistry - Anti-DDDDK tag (Binds to FLAG® tag sequence) antibody (ab1162)Image from Kapoor N et al., J Biol Chem. 2011 Apr 22;286(16):14073-9. Epub 2011 Jan 4. Fig 1A.; doi: 10.1074/jbc.M110.185298; April 22, 2011 The Journal of Biological Chemistry, 286, 14073-14079.

    ab1162 staining DDDDK tag (pink) in neonatal rat cardiomyocytes transduced with Ad-TBX18/FLAG®-IRES-GFP by Immunocytochemistry/ Immunofluorescence.

    Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton X-100 and incubated with primary antibody. An Alexa Fluor®-conjugated anti-rabbit IgG was used as the secondary antibody. Nuclei were stained with DAPI (blue).

实验方案

  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

数据表及文件

  • SDS download

  • Datasheet download

    Download

文献 (114)

发表研究结果有使用 ab1162?请让我们知道,以便我们可以引用本数据表中的参考文章。

ab1162 被引用在 114 文献中.

  • Garg M  et al. Cardiolipin-mediated PPAR? S112 phosphorylation impairs IL-10 production and inflammation resolution during bacterial pneumonia. Cell Rep 34:108736 (2021). PubMed: 33567272
  • Hall ET  et al. Cytoneme delivery of Sonic Hedgehog from ligand-producing cells requires Myosin 10 and a Dispatched-BOC/CDON co-receptor complex. Elife 10:N/A (2021). PubMed: 33570491
  • Murakami T  et al. Regulation of MRE11A by UBQLN4 leads to cisplatin resistance in patients with esophageal squamous cell carcinoma. Mol Oncol 15:1069-1087 (2021). PubMed: 33605536
  • Li N  et al. Upregulation of deubiquitinase USP7 by transcription factor FOXO6 promotes EC progression via targeting the JMJD3/CLU axis. Mol Ther Oncolytics 20:583-595 (2021). PubMed: 33768140
  • Liu B  et al. SKP2-Promoted Ubiquitination of FOXO3 Promotes the Development of Asthma. J Immunol 206:2366-2375 (2021). PubMed: 33837090
View all Publications for this product

客户评价及客户问答

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1-10 of 21 Abreviews or Q&A

Western blot abreview for Anti-DDDDK tag (Binds to FLAG® tag sequence) antibody

Average
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Mouse Cell lysate - other (neuroblastoma2 a cells)
Gel Running Conditions
Reduced Denaturing (10)
Loading amount
10 µg
Specification
neuroblastoma2 a cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 Mar 29 2017

Immunohistochemistry (Frozen sections) abreview for Anti-DDDDK tag (Binds to FLAG® tag sequence) antibody

Inconclusive
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (brain)
Permeabilization
Yes - Triton x-100
Specification
brain
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2.5% · Temperature: 25°C
Fixative
Paraformaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

久仁子 島崎

Verified customer

提交于 Sep 05 2016

Western blot abreview for Anti-DDDDK tag (Binds to FLAG® tag sequence) antibody

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (HeLa)
Gel Running Conditions
Reduced Denaturing (10%)
Loading amount
10 µg
Specification
HeLa
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 Apr 22 2016

Immunohistochemistry (PFA perfusion fixed frozen sections) abreview for Anti-DDDDK tag (Binds to FLAG® tag sequence) antibody

Inconclusive
Abreviews
Abreviews
Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Mouse Tissue sections (Brain- striatal medium spiny neurons)
Antigen retrieval step
None
Permeabilization
Yes - 0.03% Triton
Specification
Brain- striatal medium spiny neurons
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 20°C
Fixative
10% Formalin
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Rivka Schwarcz

Verified customer

提交于 Jul 28 2014

Immunocytochemistry/ Immunofluorescence abreview for Anti-DDDDK tag (Binds to FLAG® tag sequence) antibody

Average
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Monkey Cell (COS7)
Permeabilization
Yes - Tritonx 100
Specification
COS7
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
Formaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 Oct 18 2013

Immunocytochemistry/ Immunofluorescence abreview for Anti-DDDDK tag (Binds to FLAG® tag sequence) antibody

Good
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (hTERT-RPE1)
Permeabilization
Yes - saponin
Specification
hTERT-RPE1
Blocking step
BSA as blocking agent for 20 minute(s) · Concentration: 0.02% · Temperature: 18°C
Fixative
Paraformaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 Dec 19 2012

Western blot abreview for Anti-DDDDK tag (Binds to FLAG® tag sequence) antibody

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (hTERT-RPE1)
Gel Running Conditions
Reduced Denaturing (8 % acrylamid)
Loading amount
50000 cells
Treatment
transfected with flag tag plasmid 20hr
Specification
hTERT-RPE1
Blocking step
Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 Dec 14 2012

Question

Je suis à la recherche d'un anticorps anti FLAG qui fonctionnerait en
Immunogold pour la microscopie electronique,
Auriez-vous des références d'anticorps à me conseiller?
Merci d'avance

Read More

Abcam community

Verified customer

Asked on Dec 12 2012

Answer

Merci de nous avoir contactés.

Nous avons plusieurs anti-FLAG anticorps au catalogue par contre aucun n’a été testé en microscopie électronique.

Cependant, je peux vous proposer :

ab1162 (www.abcam.com/ab1162)
ab 18230 (www.abcam.com/ab18230)
ab21536 (www.abcam.com/ab21536)

Ces anticorps n’ont pas encore été testés en microscopie électronique et nous n’offrons pas d’échantillon gratuit à des fins de tests préliminaires. Cependant, si vous souhaitez tester l’un de ces anticorps en microscopie électronique, il me sera possible de vous offrir une remise. Après soumission d’une Abreview avec vos résultats, un avoir de la valeur de l'anticorps testé vous sera offert.

Étapes à suivre :

1. Nous informer que vous souhaitez tester un de nos anticorps en microscopie électronique et nous vous enverrons un code promotionnel (valable 4 mois) à utiliser après obtention et envoi de vos résultats,

2. Acheter l'anticorps,

3. Nous envoyer vos résultats (positifs ou négatifs) obtenus sous forme d'Abreview (www.abcam.com/abreviews).

4. L'envoi de vos résultats activera le code et vous donnera un avoir de la valeur de l'anticorps testé.

J'espère que ces informations vous aideront. N'hésitez pas à me contacter si vous avez besoin de plus de renseignements.

Read More

Abcam Scientific Support

回复于 Dec 12 2012

Question

Hola ,
En principio preferiríamos cambiar de anticuerpo y probar con el ab1259 (el que tiene conjugado el FITC). Como solo nos interesa para immunofluorescencia, mejor un anticuerpo exclusivo para esta aplicación. Muchas gracias por todo, da gusto un servicio técnico así.
Saludos

Read More

Abcam community

Verified customer

Asked on Jan 30 2012

Answer

Gracias por tu confirmacion.


Acabo de hacer un nuevo pedido del anticuerpo ab1259 de forma totalmente gratuita. El número de pedido es XXXX. A lo largo del día recibirás una confirmación automática del mismo, indicando el plazo de entrega y los detalles. Si no lo recibierais, o si tuvierais alguna consulta sobre este pedido, por favor, no olvidéis mencionar el número indicado.

Espero que este nuevo anticuerpo funcione perfectamente en vuestra aplicación, y en caso contrario, por favor, no dudes en volvernos a contactar. Te recuerdo que todos los productos de Abcam tienen una garantía de 6 meses, pasada la cual no es posible ofrecer reemplazos o reembolsos. Por ello, si tuvierais algún tipo de problema, te animo a que lo reportes lo más rápido posible.

Si necesitáis cualquier otra información, o tenéis mas consultas o sugerencias, por favor, no dudéis en contactarnos de nuevo.

Read More

Abcam Scientific Support

回复于 Jan 30 2012

Question


Gracias por tu rápida respuesta.

Respecto a la queja para el anticuerpo ab1162 ya he rellenado todo el formulario, igualmente te lo resumo. El anticuerpo tanto para wb como para immunoprecipitación funciona perfectamente, pero para immunofluorescencia no es específico, ya que cuando lo pruebo tanto en células sin transfectar o transfectadas con el plásmido vacío que utilicé inicialmente para clonar luego mi proteína, la señal luego en el microscopio es muy fuerte en ambos casos. Vaya, que no hay diferencia de señal entre que exprese mi proteína con el tag (flag) o no la exprese. Y claro, eso no debería ser posible ya que el tag Flag es totalmente artificial.

Te adjunto unas imágenes tomadas en células HEK293T en las condiciones que antes te describía (plásmido vacío y con inserto) y verás que no hay especificidad alguna.
En fin, gracias por todo!

Read More

Abcam community

Verified customer

Asked on Jan 23 2012

Answer

Te agradezco mucho que nos hayas facilitado mas detalles del protocolo y te hayas tomado la molestia de enviar las imágenes.

Es evidente que el anticuerpo no es especifico contra el DDDDK tag visto que da la misma señal en células transfectas y en células sin el tag unido a la proteína. Es curioso, sin embargo, que si lo sea en WB e IP. ¿Habéis utilizado el mismo control negativo en WB?

Me comentas que este problema lo tuvisteis en el pasado, en el momento de adquirir el anticuerpo, por lo que entiendo que podemos descartar que el problema se deba a la perdida de actividad debido a las condiciones de almacenamiento del mismo a lo largo del tiempo (para mantener en perfecto estado los anticuerpos durante más de un año, se recomienda guardarlos a -20C).

El protocolo parece perfecto, no creo que ninguna mejora que pueda hacerse haga obtener una señal más específica. En estos casos es importante comprobar que tanto la transfección del plásmido a la célula se ha realizado con éxito, así como la inserción del tag en la proteína diana. En vuestro caso lo habéis comprobado con otras técnicas, por lo que no cabe duda acerca de ésto.

Quizás ayudaría a mejorar el resultado intensificar las condiciones de permeabilización de las células. Solemos recomendar si se usa Saponin, una concentración al 0.5% durante unos diez minutos.





Así que quedo a la espera de que me indiques la mejor solución para vosotros. Si tienes cualquier otra consulta, por favor, no dudes en volver a contactarnos.

Read More

Abcam Scientific Support

回复于 Jan 23 2012

1-10 of 21 Abreviews or Q&A

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