Is there anyone who has done extraction of salivary gland Aedes aegypti? Please suggest or share your experience. What is the best way to extract salivary gland Aedes aegypti? I have done it but not successfully because after running the protein by SDS PAGE, the protein band is not clear. The procedure of salivary gland extraction which I have done is salivary glands were taken (100 pairs) from adult females and were dissected in PBS and PMSF, and lysis buffer was added (1.5 mM MgCl2, 10mM Tris HCl, 10 mM NaCl, 1% Nonidet P-40 and 2mM EDTA). I used a micropistile for homogeneous, and sonicated it on an ice water bath before centrifugating at 12.000 rpm for 15' at 4o C. The protein was separated by SDS PAGE using separating gel 12% and 4 % stacking gel.
I used to dissect giant salivary glands from the LARVA of fruit fly under dissecting microscope rather than the adult insect.
thank you for your comment, yes I'm agree that isolation of larva salivary gland is easier than adult and i have ever done with drosophila larva. But I mean how is the best protein extraction of salivary gland aedes, because after running the protein by SDS PAGE, the protein band is not clear.
For the best visualization on SDS, better if using other stainning methods such as Silver stain (it is more sensitive than CBB)
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