Hela-DFO treated (0.5mM, 24h) Whole Cell Lysate (ab116322)

製品の概要

  • 製品名Hela-DFO treated (0.5mM, 24h) Whole Cell Lysate
  • 特記事項We recommend aliquoting the extracts into single use fractions and then storing them at -80°C.

    Product was previously marketed under the MitoSciences sub-brand.


    The cells were treated for 24 hours with a 0.5mM solution of Deferoxamine (DFO). DFO is used as a hypoxia-mimetic agent to stabilize Hypoxia Inducible Factor 1 (HIF-1). DFO stabilizes HIF-1 through the inhibition of Prolyl Hydroxylases (PHDs) which target HIF-1 through degradation. The mechanism of DFO inhibition is likely through the chelation of Fe2+ bound to the active site of PHD which is required for enzymatic activity.

  • アプリケーション適用あり: WBmore details

製品の特性

  • 製品の状態Liquid
  • 保存方法Shipped on Dry Ice. Upon delivery aliquot. Store at -80°C. Avoid freeze / thaw cycle.
  • バッファーpH: 7.50
    Note: This lysate was normalised with SDS-PAGE sample buffer to current concentration.
  • Concentration information loading...
  • ライゼート 備考The cells were treated for 24 hours with a 0.5mM solution of Deferoxamine (DFO). DFO is used as a hypoxia-mimetic agent to stabilize Hypoxia Inducible Factor 1 (HIF-1). DFO stabilizes HIF-1 through the inhibition of Prolyl Hydroxylases (PHDs) which target HIF-1 through degradation. The mechanism of DFO inhibition is likely through the chelation of Fe2+ bound to the active site of PHD which is required for enzymatic activity.
  • 背景HeLa cells are human epithelial cells from a fatal cervical carcinoma. The cell line was derived from cervical cancer cells taken from Henrietta Lacks, in 1951. Horizontal gene transfer from human papillomavirus 18 (HPV18) to human cervical cells created the HeLa genome which is different from either parent genome in various ways including its number of chromosomes. HeLa cells have a modal chromosome number of 82, with 4 copies of chromosome 12 and 3 copies of chromosomes 6, 8, and 17. HeLa cells are adherent cells (they stick to surfaces) and maintain contact inhibition in vitro.

アプリケーション

Our Abpromise guarantee covers the use of ab116322 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
WB Use at an assay dependent concentration.

Hela-DFO treated (0.5mM, 24h) Whole Cell Lysate 画像

  • All lanes  stained with Optiblot Blue (ab119211) (20 ml)

    Lane 1 : Hela-DFO treated (0.5mM, 24h) Whole Cell Lysate (ab116322)
    Lane 2 : Hela-Vehicle treated (Negative Control) Whole Cell Lysate (ab116321)

    Lysates/proteins at 10 µg per lane.
  • All lanes : Anti-HIF-1 alpha antibody (ab103063) at 2 µg/ml

    Lane 1 : Hela-Vehicle treated (Negative Control) Whole Cell Lysate (ab116321)
    Lane 2 : Hela-DFO treated (0.5mM, 24h) Whole Cell Lysate (ab116322)

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

Hela-DFO treated (0.5mM, 24h) Whole Cell Lysate (ab116322) 使用論文

ab116322 has not yet been referenced specifically in any publications.

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I've linked the two recommended protocols regarding nuclear fractionation and WB below:

http://www.abcam.com/assets/popups/popup_datasheet_protocols_printer_friendly.cfm?pid=239&intAbId=1

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"