Application
Western blot
Sample
Mouse Cell lysate - whole cell (mouse embryonic stem cell)
Loading amount
200000 cells
Specification
mouse embryonic stem cell
Treatment
1uM RA for 24 hours
Gel Running Conditions
Reduced Denaturing (10% gel)
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5%
Other product details
Dilution
1/500
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: block
Secondary antibody
Name
Non-Abcam antibody was used: goat anti-rabbit HRP
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+
Exposure
1 minute(s) and 0 second(s)
Bands
Specific: 48 kDa Non-specific: 35 kDa
Positive control
Lane 3: WT D3 mouse ES cells cultured without LIF and with 1uM RA for 24hours.
Negative control
Lane 1: WT D3 mouse ES cells cultured with LIF.
Lane 2: WT D3 mouse ES cells cultured without LIF for 24 hours.
Lane 2: WT D3 mouse ES cells cultured without LIF for 24 hours.
Additional data
Additional Notes
The specific band at 48kDa is only observed in the cells treated with retinoic acid (RA) for 24 hours in the absence of LIF, a previously reported method for stimulating the up-regulation of Stra8.
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提交于 Nov 30 2012