Application
Western blot
Loading amount
5 µg
Gel Running Conditions
Reduced Denaturing (15%)
Sample
Human Cell lysate - whole cell (undifferentiated human iPS cells)
Specification
undifferentiated human iPS cells
Blocking step
5% milk in TBST as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Other product details
Concentration
1 µg/ml
Incubation time
16 hour(s) and 0 minute(s) · Diluent: 5% milk in TBS
Secondary antibody
Name
Non-Abcam antibody was used: Goat anti-rabbit IgG (Dako)
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/2000
Detection
Detection method
ECL
Bands
Specific: 45 kDa
Exposure
5 minute(s) and 0 second(s)
Positive control
Sample positive control. Single band approx 45kDa.
Additional data
Additional Notes
Undifferentiated human iPS cells were lysed in RIPA buffer with protease inhibitors and performed under reducing conditions (10% gel). Following block overnight the samples were probed using an anti-Oct4 antibody (ab19857) at 1ug/ml, followed by incubation with the relevant secondary antibody at 1:2000. Samples were visualised using ECL for 5 mins and a single band at approximately 45kDa was observed.
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投稿 Jan 22 2014