User:Igor R. Kuznetsov/Notebook/RBL-2H3/2010/08/03

RBL-2H3

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U937 Delta ST on VCAM-1 and Poly-L-lysine, activated (N-fMLFF) Sample 1: Coverslips base-treated, coated with Recombinant Human VCAM-1/CD106 Fc Chimera (R&D Systems #P19320) for 30 minutes at room temperature, and gently washed 2 times with Hepes buffer. Treated with 0.5g% BSA for 30 minutes. Cells resuspended in Hepes on the coverslips for 30 minutes at room temperature, then washed with Hepes to remove unattached cells. Finally, treated with N-fMLFF [math]\displaystyle{ 10^{-7} }[/math] PBS buffer. Cells are attached, but remain globular with no protrusions. Maximum cell diameter in XY plane is seen at about 2 [math]\displaystyle{ \mu m }[/math] height. No significant cytoplasmic activity. Sample 2: Coverslips coated with Poly-L-lysine solution (P4832 Sigma) for 30 minutes at room temperature, and gently washed 2 times with Hepes buffer. Treated with 0.5g% BSA for 30 minutes. Cells resuspended in Hepes on the coverslips for 30 minutes at room temperature, then washed with Hepes to remove unattached cells. Finally, treated with N-fMLFF [math]\displaystyle{ 10^{-7} }[/math] PBS buffer. Cells are attached and develop lamella. Plenty of visible cytoplasmic activity.