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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 (38)

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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

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HRP Anti-DDDDK tag (Binds to FLAG® tag sequence) antibody [M2] (ab49763)
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Anti-V5 tag antibody (ab9137)

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.

    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

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

The Abpromise guarantee

Abpromise保証は、 次のテスト済みアプリケーションにおけるab1257の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション Abreviews 特記事項
ELISA
1/100 - 1/500.
WB (4)
1/1000 - 1/30000.
ICC/IF (2)
1/200 - 1/2000.
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

  • SDS download

  • Datasheet download

    Download

参考文献 (38)

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 38 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
  • Springgay LK  et al. Rhesus Macaque Rhadinovirus Encodes a Viral Interferon Regulatory Factor To Disrupt Promyelocytic Leukemia Nuclear Bodies and Antagonize Type I Interferon Signaling. J Virol 93:N/A (2019). PubMed: 30626678
  • 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
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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