Products:Tags & Cell Markers >> Subcellular Markers >> Cytoskeleton >> Microtubules
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ab11309 staining beta Tubulin in Mouse cultured cortical neurons by Immunocytochemistry/Immunoflourescence. Cells were plated on a poly-L-Lysine coated cover slip before being washed twice with PBS and fixed with 4% PFA at room temperature. Cells were permeabilized in 0.5%(w/v) saponin in PBS prior to blocking in 10% fetal calf serum in PBS-saponin for 2 hours at 23ºC. The primary antibody was diluted 1/1000 in the blocking buffer and incubated with the sample for 2 hours at 23ºC.
After additional washing cover splips were mounted on glass slides with ProLong® Gold antifade reagent with DAPI (Invitrogen). The image was acquired with a 40x objective.
This image is courtesy of an anonymous Abreview.
All lanes : Anti-beta Tubulin antibody [TUB 2.1] (Cy3 ®) (ab11309) at 1/1000 dilution
Lane 1 : HEK cell lysate at 5 µg
Lane 2 : HEK cell lysate at 40 µg
Performed under reducing conditions.
Predicted band size : 50 kDa
Observed band size : 55 kDa (why is the actual band size different from the predicted?)
Exposure time : 2 minutes
Lysis Buffer was 1% Triton X-100, 20mM TrisHCl pH 7.5, 10mM EGTA, 150mM NaCl, protease inhibitors. The image was acquired by scanning the blot with a FLA-9000 Starion laserscanner at 300V and 200µm resolution. The above mentioned exposure time is just the time it took to scan the blot but does not correlate to signal intensity.
This image is courtesy of an anonymous Abreview.
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