Hi,
We are studying the retina in a knockout mouse model. Compared with the control wild-type retinal lysate, the knockout mouse retinal lysate often looks cloudy. We lyze the mouse retinas in standard tissue lysis buffer (0.5% triton X-100) by a hand homogenizer at room temperature and incubate the lysate at 4 degree for half to one hour. The lysate appears cloudy after the incubation, but we can't get it clear after centrifugation at max speed for 10 mins for several rounds. We think this cloudy appearance may be associated with the gene knockout. I wonder whether somebody knows what can cause the cloudy appearance. Is it possible that abnormal lipid composition can cause it? Thanks.
Hey Jun,
since it only apperas in the KO retina samples, it is very likley that it is due to a effect cause by the gene deletion. However, if you like you can try the method for retinal tissue lysation we used. We sonicated the retinal tissue in ice cold PBS (with phosphatase and protease, for Western Blot analysis).
For the protocol you can ask Anne Wolf (here on research gate) and have a look at our publication, if you like:
Article The TSPO-NOX1 axis controls phagocyte-triggered pathological...
All the best and stay healthy,
Marc
