Salivary glands extract (SGE) preparation
Salivary glands were dissected from 100 adult cold anesthetized An. culicifacies female mosquitoes
(SF and BF each) under stereomicroscope (4X magnification) using fine needles and
pooled in Phosphate Buffered Saline (PBS; 10mM Na2SO4, 145mM NaCl (pH 7.2) containing
protease inhibitors (Complete, Roche Diagnostics, Germany). 100 pairs of pooled salivary
glandswere homogenised by ultrasonication (3 pulses of 20 sec each) on ice. Salivary gland
homogenate suspension was then centrifuged for 10 min at 5000 rpm at 4°C. Cell debris pellet
was discarded and supernatant containing salivary gland protein extract (SGE) was stored at
-80°C until further investigations. Protein concentration in the SGEs was quantified by Lowry’s
method (GeNeiTM Protein Estimation Kit) using BSA as a standard [22].