Application
Western blot
Loading amount
5 µg
Gel Running Conditions
Reduced Denaturing (10%)
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
1 hour(s) and 0 minute(s) · Diluent: 5% milk in TBS
Secondary antibody
Name
Non-Abcam antibody was used: Dako goat anti-rabbit IgG
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: 50 kDa Non-specific: 60, 39 kDa
Exposure
5 minute(s) and 0 second(s)
Negative control
50kDa (strong), 60kDa (strong) and 39kDa (weak)
Positive control
Human iPS cells - undifferentiated. There are always some cells that are undergoing differentiation at any given time and thus cells expressed early progenitor markers. Specific band observed alongside some non specific ones.
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 FoxG1 antibody (ab18259) at 1ug/ml, followed by incubation with the relevant secondary antibody at 1:2000. Samples were visualised using ECL for 5 mins and a specific band at approximately 50kDa was observed. However, a strong non-specific band was also observed at 60kDA and a weak one at 39kDa. Although undifferentiated cells may not express FoxG1, there are obviously some progenitor cells within the sample that have begun differentiation.
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投稿 Feb 24 2014