Application
Western blot
Sample
Human Cell lysate - whole cell (neurons from iPS cells)
Gel Running Conditions
Non-reduced Denaturing (12%)
Loading amount
20 µg
Specification
neurons from iPS cells
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: RT°C
Other product details
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 5% milk in TBST
Dilution
1/20000
Secondary antibody
Dilution
1/2000
Name
Non-Abcam antibody was used: goat anti rabbit IgG HRP (dako)
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Detection
Detection method
ECL+
Exposure
1 second(s)
Bands
Specific: 34 kDa
Additional data
Additional Notes
Human stem cell derived neurons were cultured for 30 days prior to collection and lysed in RIPA buffer with protease inhibitors. Proteins were resolved under reducing conditions (12% gel). Following block (5% milk in TBST), the samples were probed using an anti synaptophysin antibody at 1:20000 in blocking buffer overnight at 4C. This was followed by incubation with the relevant secondary antibody at 1:2000. Samples were visualised using ECL+ for 1s and a specific band at 34kDa observed.
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Submitted Jan 15 2016