Proximity selection using an existing antibody

Proximity seiectlon using a guide antibody on an adherent cell line 

Grow the cell line under normal culture conditions on chamber slides to approx. 80% confluence (approx. 1 X 10 to 1 x 10 cells per well). Cells can either be left unfixed or fixed with a variety of fixation procedures, for example treatment with 0.1% glutaraldehyde for 15 min at room temperature. If unfixed cells are used, carry out all the following steps at 4 °C. 

Block the cells in 100 p,l PBS% containing 3% skimmed milk for 1-2 h at room temperature. 

Gently wash the cells with three changes of PBS. 

Add the primary guide antibody at an appropriate concentration in PBS containing 3% skimmed milk. Incubate for 1 h at room temperature. 

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