Application
Western blot
Sample
Human Cell lysate - whole cell (HMEC, MCF-10A, LN229, MDA-MB231 cell lines)
Loading amount
10 µg
Specification
HMEC, MCF-10A, LN229, MDA-MB231 cell lines
Gel Running Conditions
Reduced Denaturing (10% SDS-PAGE)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Other product details
Dilution
1/500
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: BLOCKING BUFFER
Secondary antibody
Name
Non-Abcam antibody was used: 926-32223
Host species: Donkey
Clonality: Polyclonal
Conjugation: IRDye® 800CW
Host species: Donkey
Clonality: Polyclonal
Conjugation: IRDye® 800CW
Dilution
1/1000
Detection
Detection method
LI-COR odyssey
Exposure
5 minute(s) and 0 second(s)
Bands
Specific: 52 kDa Non-specific: 64 kDa
Negative control
LANE 1:HMECs(human mammary epithelial cell line).
LANE 2:MCF-10A (human mammary epithelial cells immortalized)
LANE 3:LN229 (glyoblastoma cell line)
LANE 4: MDA-MB231 (transformed mammary epithelial cells)
LANE 2:MCF-10A (human mammary epithelial cells immortalized)
LANE 3:LN229 (glyoblastoma cell line)
LANE 4: MDA-MB231 (transformed mammary epithelial cells)
Additional data
Additional Notes
I obtained a strong single band at 52 kDa but the predicted molecular weight is 59 kDa. Also, in lane 4 I've got a another faint band at 64 kDa.
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.
Ms. barbara chaneton
Verified customer
投稿 Jan 25 2011