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Senescent-associated b-Gal Assay

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929

 

  1. Wash cells with PBS
  2. Fix cells in 0.2% glutaraldehyde for 5’
  • dilute this in PBS
    1. Wash cell with PBS
    2. Stain with X-gal solution (ingredients listed below) for 6 to 24 hours at 37 °C

    Chemical Conc.

    Stocks Amount needed for 10 plates (40 ml)
    1 mg/ml X-gal 40 mg/ml  1 ml
    150 mM NaCl 5 M 1.2 ml
    2 mM MgCl2 1M 80 μ l
    5 mM K3 Fe(CN)6 100 mM  2 ml
    5 mM K4 Fe(CN)6 100 mM 2 ml
    40 mM NaPi, pH 6.0  0.1M 16 ml

    17.72 ml dd H2 O

    NaPi - 0.1M at pH 6.0

    • 2.5 ml Na2 HPO4
    • 17.6 ml NaH2 PO4
    • dd H2 O to 200 ml

     

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