PreparationMethod

HeLa cells are cultured using standard media condition until semi-confluent (70-80%).The cells are grown under true hypoxic conditions where the oxygen level in the incubator is reduced to 2% for 4 hours to accumulate Hif-1a within the cells.The cells are then washed in PBS and lysed into 1X Laemmli sample buffer with BME.The lysate is sonicated and tested by Western blot for reactivity to Hif-1a using a polyclonal anti-Hif-1a antibody (NB100-134).Tubulin reactivity of each lysate is shown as a loading control.

Dilutions	SDS-Page 10 ul / lane
Application Notes	HeLa Whole cell lysate is provided as a Western blot positive control for Hif-1 research. The lysates are provided as a set of treated and untreated samples.Use 10 ul per lane for a standard mini-gel blot (approx. 1 mg/ml).

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