NIH 3T3 Hypoxic / Normoxic Cell Lysate

SDS-PAGE

SDS-Page (10 ul / lane)

NIH 3T3 whole cell lysate is provided as a Western blot positive control. The lysates are provided as a set of treated and untreated samples. Use 10 ul per lane for a standard mini-gel blot (1.0 mg/ml).

Cell

Hypoxic / Normoxic

NIH 3T3 cells are grown under standard tissue culture conditions until semi-confluent (70-80%). The cells are either grown under normoxic conditions or true hypoxic conditions where the oxygen level in the incubator is reduced to 2% for 4 hours. The cells are then washed in PBS and directly lysed into 1X Laemmli sample buffer with BME. The lysate is sonicated and tested by Western blot.

The protein lysate is prepared in 1x Laemmli sample buffer with BME. Boiling before loading is not necessary.

1.0 mg/ml

Store at -20 to -80C. Avoid freeze-thaw cycles.