The banding was visualized at short UV light Carozzi et al., 1991.
Where a band was visualized at the correct expected size for either exoA or oprL loci, the specimen was considered positive for P. aeruginosa.
Bands were visualized by gel electrophoresis (neo band at ∼270 bp and WT band at 108 bp).
The membranes were washed 3 times in TBST and incubated with HRP-conjugated secondary antibody (Chemicon International) at a dilution of 1∶2500 for 1 hour at RT. Immunoreactive bands were visualized using the enhanced chemiluminescence method (GE Healthcare, Buckinghamshire, UK).
As shown previously, three protein bands were visualized with anti- srcFK antibodies: a doublet at 59/60 kDa and another at ∼54 kDa, corresponding to the src-family members src, fyn, and yes, respectively.
Bands were visualized by gel electrophoresis (DA transporter band at 565 bp and eGFP band at ∼300 bp).
Following three washes with Tris-buffered saline, the protein bands were visualized with LuminGLO for 1 min at room temperature.
For gangliosides, the HPTLC plates were developed with CHCl3: CH3OH: 0.02% calcium chloride (55:45:10 by volume), and the bands were visualized with resorcinol spray and burning at 100℃ for 10 min (Hauser et al., 2004; Kasperzyk et al., 2004).
The membranes were then washed and incubated with the appropriate peroxidase conjugated secondary antibodies for 1 h at RT. Immunoreactive bands were visualized by ECL GE Healthcare Amershamm) and the bands were quantified by densitometry with NIH Image J software.
After extensive washing in Tris-buffered saline containing 0.1% Tween 20 at room temperature, bands were visualized by the enhanced chemiluminescence method (Roche Molecular Biochemicals).
