I want to extract the midgut of Callosobruchus maculatus larvae to isolate the protease. This will be my first time dissecting a very minute larvae (2nd/3rd instar). I read some publications about how they dissect and prepare the gut homogenate. However, some research protocols were different with the others. Sometimes, the insects were dissected by only using cold distilized water, but others by using buffers (citrate-phosphate or sodium-acetate) or NaCl solution.
Dear Ito,
For isolating and collecting the fat body from Drosophila larvae we used 1X PBS as the fat body tissue disintegrated in water
Good luck
Use phosphate-buffered saline (PBS) solution: [ 81 mM Na2HPO4, 19 mM Na2HPO4, 150 mM NaCl, pH 7.4). My students before used this to extract the midgut of Aedes aegypti fourth instar larvae. It worked since they managed to isolate midgut bacteria under aseptic conditions.
Mayer Calma Kindly look at component of PBS. You mentioned Na2HPO4 twice.
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