Application
Western blot
Sample
Human Cell lysate - whole cell (U87-MG, human brain, lymphoblastoid cell-line)
Loading amount
15 µg
Specification
U87-MG, human brain, lymphoblastoid cell-line
Gel Running Conditions
Reduced Denaturing (4-12 % Bis-Tris Gel)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Other product details
Dilution
1/400
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 5 % Milk in TBS-T
Secondary antibody
Name
Non-Abcam antibody was used: SC-2004 Goat anti-rabbit IgG-HRP
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/7500
Detection
Detection method
ECL
Exposure
1 minute(s) and 0 second(s)
Bands
Specific: 45-48, 55-60 kDa Non-specific: 38 kDa
Positive control
U87-MG-lysate, human brain-lysates
Additional data
Additional Notes
Nice clean bands, the 48 kDa-band seems to be specific as it occurs in all the sample. However we`re unsure about the band below 60 kDa occuring in the brain lysates. It could be due to phosphorylation.
Abcam response
It band at 60kDa might be the glycosylated form of the protein; the santa cruz antibody sc-74497 shows similar pattern;
protocol OK, image OK. PS 11.1.2012
protocol OK, image OK. PS 11.1.2012
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. Georg Ziegler
Verified customer
投稿 Jan 11 2012