Application
Western blot
Loading amount
100 µg
Gel Running Conditions
Reduced Denaturing (8 %)
Sample
Human Cell lysate - whole cell (NB4 and K562 human cell lines)
Specification
NB4 and K562 human cell lines
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Other product details
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 1 % Milk in PBS-T buffer
Dilution
1/5000
Secondary antibody
Name
Non-Abcam antibody was used: Pierce Goat Anti-Rabbit Poly-HRP
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/5000
Detection
Detection method
ECL
Bands
Specific: 130, 125 kDa Non-specific: 180, 80, 53, 40, 30 kDa
Exposure
1 minute(s) and 0 second(s)
Additional data
Additional Notes
PML-RARa should be around 110 kDa, but appears at 130 kDa in the western blot of NB4 cell lysates (A and B). The K562 cells in fig. B were used to determine if the 130 kDa protein band is specific, because K562 cells do not contain the PML-RARa protein.
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.
MR. Christian Marx
Verified customer
提交于 Jul 31 2013