1. ECM's are stored and diluted as follows:

-rat tail collagen, type I (3.66 mgs/ml), stored in claire's 4*, stable for 9 months, opened 5/1/93. Dilute for coating in 0.02N acetic acid.

-mouse laminin (1.11 mgs/ml), stored in claire's -70* in 90ul aliquots, stable for 9 months, opened 5/1/93. Dilute for coating in serum free media.

-human fibronectin (lyophilized powder), stored in claire's 4*, stable for 18 months, opened 5/1/93.

Solubilize in sterile dH2O at a concentration of 1mg/ml, allow 30 min to go into solution.

Sterile filter, aliquot, quick freeze and store at -70*. Stable for 2 weeks. Dilute for coating in serum free media.

2. In a plastic 8 well chamber slide, add 1.25ug ECM to each well in appropriate dilution media (see above). Each well will hold 500ul volume total.

3. Incubate at room temperature for 1 hour.

4. Rinse each well with tissue culture PBS. [ASPIRATE CAREFULLY, DO NOT SCRAP BOTTOM].

5. Slides can be stored at this point at 4* damp or air dried, if sterility is maintained.

6. Prior to plating cells in wells, block wells with 1% BSA in tissue culture PBS for 1 hour at room temp. or overnight at 4*.