thermal energy turns on the gene (heat up the cells, not too much to denature)

To get rid of the medium, and "fix" the cell:
- this increases permeability of the cell, killing the cell
-paraformaldehyde fixes the structures in place, instead of rinsing away. Keeps the cells in place (HeLa sticks to the bottom of the plate very well.)

Blocks off unwanted receptor sites, so that when antibodies are added, they attach to the correct receptor site. 

Ab's fit onto the appropriate receptor sites 

Secondary blocker blocks off unwanted receptor sites so that when the second antibodies are added, they attach to the primary Ab instead of confounding receptor sites. 

Secondary Ab attaches onto Primary Ab, Secondary Ab is a flourophore, carries the part that glows. 
Lots of flourophores bind to one primary antibody, so you have a lot of signal with having multiple bindings. 

Seeing how brightly it glows to see how much heat shock protein is present compared between the two cell samples. 
looking at how bright the thing is to see how much concentration of the HSP there is present. 
Is there an elevated concentration of the HSP

To get a visual of our results. 

-thermal energy "turns on" the gene (like chemistry in lac operon)
-Using HeLa Cells

How the blockers work: 
- Blocker 1, Blocker 2
- Block off extra receptor site that the antibody could attach to 
-Blocks off confounding sites, and the blockers stick. 

Washing more thorough than rinsing
- add solution
-wait x minutes
-wash/rinse extra away
      -rinsing: with Phosphate Buffered Saline (PBS)

-Take an average brightness of 4-5 cells from both samples
-T-Test: whether there really is a significant difference in brightness (yes there is a difference, no there isn't a difference)