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  1. Link

    ddddk-tag-binds-to-flag-tag-sequence-antibody-ab1257.pdf

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Tags & Cell Markers Epitope Tags DDDDK Tag
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Anti-DDDDK tag (Binds to FLAG® tag sequence) 抗体 (ab1257)

  • Datasheet
  • SDS
Reviews (8)Q&A (2)References (37)

Product price, shipping and contact information

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出荷および受注について

Abpromise

保証された製品品質、優れたカスタマー・サポート。 詳細はこちら。

Immunocytochemistry/ Immunofluorescence - Anti-DDDDK tag (Binds to FLAG® tag sequence) antibody (ab1257)
  • Western blot - Anti-DDDDK tag (Binds to FLAG® tag sequence) antibody (ab1257)

Key features and details

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

こちらの製品もご検討ください

二次抗体
Product image
Donkey Anti-Goat IgG H&L (HRP) (ab97110)

関連製品

製品の概要

  • 製品名

    Anti-DDDDK tag (Binds to FLAG® tag sequence) antibody
    DDDDK tag (Binds to FLAG® tag sequence) 一次抗体 製品一覧
  • 製品の詳細

    Goat polyclonal to DDDDK tag (Binds to FLAG® tag sequence)
  • 由来種

    Goat
  • アプリケーション

    適用あり: ELISA, WB, ICC/IFmore details
  • 種交差性

    交差種: Species independent
  • 免疫原

    Synthetic peptide corresponding to DDDDK tag conjugated to Keyhole Limpet Haemocyanin (KLH).
    Sequence:

    xxxDDDDK

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
  • 特記事項

    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.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    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, as well as customer reviews and Q&As.

製品の特性

  • 製品の状態

    Liquid
  • 保存方法

    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • バッファー

    pH: 6.8
    Preservative: 0.1% Sodium azide
  • Concentration information loading...
  • 精製度

    Immunogen affinity purified
  • ポリ/モノ

    ポリクローナル
  • アイソタイプ

    IgG
  • 研究分野

    • Tags & Cell Markers
    • Epitope Tags
    • DDDDK Tag

関連製品

  • Compatible Secondaries

    • Donkey Anti-Goat IgG H&L (Alexa Fluor® 488) (ab150129)
    • Donkey Anti-Goat IgG H&L (HRP) (ab205723)
  • Isotype control

    • Goat IgG, polyclonal - Isotype Control (ab37373)

アプリケーション

Our Abpromise guarantee covers the use of ab1257 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
ELISA 1/100 - 1/500.
WB 1/1000 - 1/30000.
ICC/IF 1/200 - 1/2000.

ターゲット情報

  • 関連性

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

    • 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

画像

  • Immunocytochemistry/ Immunofluorescence - Anti-DDDDK tag (Binds to FLAG® tag sequence) antibody (ab1257)
    Immunocytochemistry/ Immunofluorescence - Anti-DDDDK tag (Binds to FLAG® tag sequence) antibody (ab1257)

    Human microvascular endothelial cells expressing DDDDK-tagged beta-catenin following transient transfection.
    Ab1257 used at 1 µg/ml. Detected with FITC labeled rabbit anti-goat IgG (H&L) secondary antibody.

  • Western blot - Anti-DDDDK tag (Binds to FLAG® tag sequence) antibody (ab1257)
    Western blot - Anti-DDDDK tag (Binds to FLAG® tag sequence) antibody (ab1257)
    All lanes : Anti-DDDDK tag (Binds to FLAG® tag sequence) antibody (ab1257) at 0.04 µg/ml

    Lane 1 : E. coli whole cell lysate. at 0.2 µg
    Lane 2 : E. coli whole cell lysate. at 0.1 µg
    Lane 3 : E. coli whole cell lysate. at 0.05 µg


    Western Blot analysis of E. coli Whole Cell Lysates, labelled with DDDDK-tagged Protein by ab1257 at 1/25,000.

プロトコール

  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

データシートおよび資料

    • Datasheet
    • SDS
  • 参考文献 (37)

    ab1257 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

    ab1257 は 37 報の論文で使用されています。

    • Cui L  et al. The Lysosomal Membrane Protein Lamp2 Alleviates Lysosomal Cell Death by Promoting Autophagic Flux in Ischemic Cardiomyocytes. Front Cell Dev Biol 8:31 (2020). PubMed: 32117965
    • Busch JD  et al. MitoRibo-Tag Mice Provide a Tool for In Vivo Studies of Mitoribosome Composition. Cell Rep 29:1728-1738.e9 (2019). PubMed: 31693908
    • Xu W  et al. Endothelial FAM3A positively regulates post-ischaemic angiogenesis. EBioMedicine 43:32-42 (2019). PubMed: 31000420
    • Yin G  et al. miR-221-3p promotes the cell growth of non-small cell lung cancer by targeting p27. Mol Med Rep 20:604-612 (2019). PubMed: 31180541
    • Arrant AE  et al. Impaired ß-glucocerebrosidase activity and processing in frontotemporal dementia due to progranulin mutations. Acta Neuropathol Commun 7:218 (2019). PubMed: 31870439
    View all Publications for this product

    レビューと Q&A

    Show All レビュー Q&A
    レビューを送る 質問を送る

    1-10 of 10 Abreviews or Q&A

    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 - nuclear (Vascular smooth muscle cells)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    20 µg
    Specification
    Vascular smooth muscle cells
    Blocking step
    Milk as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
    Read More

    Abcam user community

    Verified customer

    投稿 Mar 22 2018

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-DDDDK tag (Binds to FLAG® tag sequence) antibody

    Average
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Brain)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: citrate buffer
    Permeabilization
    No
    Specification
    Brain
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 25°C
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    投稿 Mar 05 2018

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

    Below Average
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HEK293)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    20 µg
    Specification
    HEK293
    Blocking step
    Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    投稿 Feb 28 2018

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

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Rat Cell lysate - other (IP from Neonatal Rat Ventricular Myocytes infected)
    Gel Running Conditions
    Reduced Denaturing (10% SDS PAGE)
    Loading amount
    1.5 µg
    Specification
    IP from Neonatal Rat Ventricular Myocytes infected
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Dr. Chris Tracy

    Verified customer

    投稿 Dec 16 2014

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

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Mouse Tissue sections (Muscle)
    Permeabilization
    Yes - Triton 0.5%
    Specification
    Muscle
    Blocking step
    BSA as blocking agent for 45 minute(s) · Concentration: 5% · Temperature: 20°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    投稿 Oct 27 2014

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

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (Vascular smooth muscle cells)
    Permeabilization
    Yes - NP40
    Specification
    Vascular smooth muscle cells
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    投稿 Dec 16 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 (COS7)
    Permeabilization
    Yes - triton X100
    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

    Abcam user community

    Verified customer

    投稿 Oct 18 2013

    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 (Kidney cancer cell)
    Gel Running Conditions
    Non-reduced Denaturing (10% gel)
    Loading amount
    20 µg
    Specification
    Kidney cancer cell
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
    Read More

    Abcam user community

    Verified customer

    投稿 Nov 12 2010

    Question

    I would like to detect FLAG (DDDDK) by immunohistology and wanted to know if you have an antibody that you would recommend for paraffin fixed sections. We work on bone so frozen sections are a bit of a nightmare for us.

    Read More

    Abcam community

    Verified customer

    Asked on Jan 23 2006

    Answer

    Thank you for your enquiry. None of our FLAG tag antibodies have currently been tested in IHC on paraffin sections. Some have been tested on frozen sections (ab1257,ab21536) and as this work in this application this indicates that those antibodies have the ability to recognise the native form of the protein so have good chances of working in other immunohistochemical methods. They therefore have a chance of working in IHC-P, providing the fixation of the tissue does not modify the epitope dramatically and prevent recognition by the antibodies. I hope this information will help you, please do not hesitate to contact me again if you need more advice,

    Read More

    Abcam Scientific Support

    Answered on Jan 24 2006

    Question

    DESCRIPTION OF THE PROBLEM High background, multiple bands and wrong band size SAMPLE mouse stomach sonicated in RIPA buffer plus complete protease inhibitors (Roche # 1697498) on ice. The whole tissue extract was separated on 15% SDS-PAGE and thansfered to PVDF membrane. PRIMARY ANTIBODY Ab1257 goat anti-ddddk 1:1000, 1: 2000, 1: 5000, 1:10000 and 1: 15000 in PBS-T with/without 5% non-fat milk incubated for 1 hr/RT wash in PBS-T 10 minx3 time SECONDARY ANTIBODY Sigma rabbit anti-goat, HRP conjugated 1:2000, 1: 5000 in PBS-T incubated for 30 min to 1 hr/RT wash in PBS-T 10 minx3 times DETECTION METHOD ECL POSITIVE AND NEGATIVE CONTROLS USED Negative control: stomach extract without my transgene, HeLa cell extract Positive control: HeLa cell with my transgene ANTIBODY STORAGE CONDITIONS 4 C cold room in original package SAMPLE PREPARATION RIPA buffer plus complete protease inhibitors (Roche # 1697498). AMOUNT OF PROTEIN LOADED 20 ug/well ELECTROPHORESIS/GEL CONDITIONS SDS-PAGE, reducing, 15% precast gel from BioRad TRANSFER AND BLOCKING CONDITIONS Stander semi-dry transfer buffer Blocking in 5% non-fat milk HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? >10 HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? primary Ab dilution, secondary Ab dilution ADDITIONAL NOTES I used Sigma M2 Ab (monoclonal from mouse) as golden standard for FLAG tag detection. Now I turned to your product because it's cheaper. In the whole series of my western, The M2 Ab gives me a sharp, solid single band without background, which means there is no degradation in my sample.

    Read More

    Abcam community

    Verified customer

    Asked on Feb 12 2004

    Answer

    Results such as these are commonly solved by loading less lysate (try 5 mcg/lane), diluting the primary antibody more (try 1:25,000) and diluting the secondary antibody more (try 1:20,000). The goat anti-ECS is 10 to 20 fold higher titered than the Sigma M2 antibody. You could use the Sigma M2 antibody at 1:25 and would likely get results similar to those you are seeing with the goat anti-ECS.

    Read More

    Abcam Scientific Support

    Answered on Feb 23 2004

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

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