- Motility assay (w/lipids)
- Koch was unsure about the "temperature gradient" i had reported on yesterday, and I may have made a mistake when starting as such
- What happened is I got two statements mixed up
- To quote the manual:
- "Depending on various conditions within the chamber, such as flask placement and size, the heat produced by growing organisms, heat losses due to liquid evaporation from flasks, etc., the display temperature may differ from the temperatures within the flasks themselves"
- "The temperature probe measures the temperature of the air at the probe's location, near the heat exchanger return vent."
- Andy wants to try to mix up a new batch of kinesin and retry his lipids assay
- Oops, he's using the old stuff
- He also wants to redo a "standard" motility assay to make sure everything is working
- Andy has done initial testing to show that the kinesin we have and the MT's are still in good order
- The idea now is that the lipids are too concentrated (and the layers are too thick) so we will dilute them down
- Andy diluted the lipids from 10mg/ml to 1mg/ml using chloroform
- Andy spun-coated a cover glass with 1/2 lipids 1/2 naked
- Added kinesin + brb solution to the lipids and letting it sit
- Visualized the sample and there are MT's floating around, but none seem to really be sticking to the lipids
- The antifade was not working very well (or Andy forgot to add it) so most of the MT's faded quickly
- Also, the lipid bi-layer did not look very uniform
- Andy wanted to mix up some Casein in BRB80, but the BRB was not going into solution
- He told me that there is a procedure, entailing the gradual mixing of casein into the BRB. I tried mixing 1/3 of the amount we wanted and it still did not go into soln
- Andy wanted me to look up some stuff about Micelles
- In particular, size of casein micelles and how to make them
- Found a paper all about Casein Micelles by googling "Casein Micelles"
- They state that:
- "The shape of casein micelles as observed by electron microscopy is spherical, ranging in size from 50-500 nm in diameter (average about 120nm) and a molecular mass from 106-109 Da"
- Its also interesting to note that they say: "size distribution of the micelles is governed by the amount if available κ-caseins"