Western Blot: GLUT10 Antibody [NB100-490] - NCI-H1299 non-small cell lung tumor cells were transfected with the eukaryotic expression vectors pcDNA3.1/V5 (lanes 1,2), pCMV-GLUT10 (lanes 3,4), and pCMV-GLUT1 (lanes 5,6). GLUT10 was detetced using NB100-490.
Immunocytochemistry/Immunofluorescence: GLUT10 Antibody [NB100-490] - Dissected inner ear tissues from mice immersed in 20% sucrose PBS solution (pH7.4) followed by being embedded in OCT. Tissues snap frozen in liquid nitrogen & 7um sections were cut by cryostat. Cochlear sections from organ of Corti were dissected & rinsed by 0.1% Triton in PBS (pH7.4,PBST) for 30min. Sections were blocked with 5% goat serum in PBST at RT 1hr. NB100-490 was used at 1:200 to label the cochlear sections at 4C. Sections were incubated with secondary (anti-rabbit Cy3, 1:400) RT 1 hr. Slides mounted with fluoromont G and examined under a confocal microscope. Strong immunoreactivity (red) is observed in the inner and outer hair cells and basilar membrane. Spiral ganglion neurons display moderate staining.
Western Blot: GLUT10 Antibody [NB100-490] - Cochlea from embryonic mice and mice up to 1 yr old were removed and the pure membrane fraction loaded on 4-20% SDS gel. Following electrophoresis, transfer and blocking, the blot was incubated overnight at 4C with NB100-490 at 1:500 dilution followed by incubation with secondary Ab at 1:3,000 dilution for 1h at room temp. Protein bands were visualized on x-ray film with SuperSignal west femto ECL substrate. Lane 1:10ug of protein from liver Lane 2:10ug of protein from cochlea (E18) Lane 3:10ug of protein from cochlea (P7) Lane 4:10ug of protein from cochlea (P10) Lane 5:10ug of protein from cochlea (P14) Lane 6:10ug of protein from cochlea (P17) Lane 7:10ug of protein from cochlea (P60) Lane 8:10ug of protein from cochlea (1 Year)