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

Overview

  • Product nameHela-DFO treated (0.5mM, 24h) Whole Cell Lysate
  • General notesWe 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.

  • Tested applicationsSuitable for: WBmore details

Properties

  • FormLiquid
  • Storage instructionsShipped on Dry Ice. Upon delivery aliquot. Store at -80°C. Avoid freeze / thaw cycle.
  • Storage bufferpH: 7.50
    Note: This lysate was normalised with SDS-PAGE sample buffer to current concentration.
  • Concentration information loading...
  • Lysate notesThe 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.
  • BackgroundHeLa 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.

Applications

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.

Application Abreviews Notes
WB Use at an assay dependent concentration.

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

  • 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.

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

ab116322 has not yet been referenced specifically in any publications.

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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"