Application
Western blot
Sample
Human Cell lysate - whole cell (undifferentiated Human ES cells on MEFs)
Gel Running Conditions
Reduced Denaturing (12.5)
Loading amount
20 µg
Specification
undifferentiated Human ES cells on MEFs
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: RT°C
Other product details
Incubation time
12 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 5% milk in TBS
Concentration
1 µg/ml
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 minute(s) and 0 second(s)
Bands
Specific: 90 kDa
Additional data
Additional Notes
Undifferentiated human ES cells were lysed in RIPA buffer with protease inhibitors and performed under reducing conditions (12.5% gel). Following block, the samples were probed using an anti-transferrin antibody at 1ug/ml, followed by incubation with the relevant secondary antibody at 1:2000. Samples were visualised using ECL for 1 mins and a single band (weak) at approx. 90kDa was observed.
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投稿 Jan 15 2016