Application
Western blot
Sample
Rat Cell lysate - whole cell (C6 Cell line)
Loading amount
25 µg
Specification
C6 Cell line
Gel Running Conditions
Reduced Denaturing (6% gel)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Other product details
Dilution
1/1000
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C
Secondary antibody
Name
Non-Abcam antibody was used: anti-rabbit antibody
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
5 second(s)
Bands
Specific: 150 kDa
Additional data
Additional Notes
Use Non-treated C6 Cell Lysate . Load 25 micrograms total protein. Gel: 6% gel (homemade). SDS-PAGE condition: 80 volts for about 3 hours. Transfer condition: 80 volts for 120 mins. membrane: PVDF (Bio-rad 162-0177) block in 5% milk in TBST for 1 hour. primary antibody 1:1000 4 degree over night. Secondary antibody ( Dako P0448) 1:2000 in block buffer for 1 hour at RT. ECL kit (pierce 32106). Exposure time: 5 seconds.
I did not expect the molecular weight in rat cells is around 150 kD. But as shown in the figure, there was only one clear band, I really don't know how to explain it.
I did not expect the molecular weight in rat cells is around 150 kD. But as shown in the figure, there was only one clear band, I really don't know how to explain it.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.
Abcam user community
Verified customer
Submitted Jun 11 2008