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Subsequently, the blots proceeded through a second round of immunodetection by agitating for 45 min at 50C in stripping buffer (2% SDS, 0.8% -mercaptoethanol (ME), and 62.5 mM Tris-HCl, pH 6.8) 32,56,62 followed by re-probing with HRP-labeled anti-HGF N -terminal domain mAb (1:2500 dilution in CanGetSignal solution 1) to monitor HGF -chain amount on the blots as loading controls

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Notably, in female Sprague-Dawley rats, a hypoxic challenge (6% O for 1 h) followed by 20 min of reoxygenation resulted in increased BBB permeability to ( 14 C)-sucrose (~340 Da), while the permeability to Evans blue dye pre-bound to albumin (~80 kDa) remained unchanged, suggesting selective permeability changes favoring small molecules over macromolecules during acute hypoxia [84, 89, 90]

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There may be minimal negative impacts if regular checkups are done and administered properly