Application
Western blot
Sample
Mouse Tissue lysate - whole (Spinal Cord)
Gel Running Conditions
Reduced Denaturing (10% gel)
Loading amount
80 µg
Specification
Spinal Cord
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Other product details
Incubation time
18 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 5% Milk in .1M TBS and .1% Tween
Dilution
1/500
Secondary antibody
Dilution
1/12000
Name
Non-Abcam antibody was used: Licor 800
Host species: Goat
Clonality: Polyclonal
Conjugation: IRDye® 800CW
Host species: Goat
Clonality: Polyclonal
Conjugation: IRDye® 800CW
Detection
Detection method
Infrared
Negative control
LOXP S1P1 knockout mice were used with an NES Cre driver. Studies with the same tissue were performed and showed no activity with S1P1 specific agonists, namely phospho-FTY and SEW. A band was found in spinal cord from these mice (Lane 3) at the same level as wild type (Lane 2) suggesting this antibody is not specific for S1P1.
Exposure
6 minute(s) and 0 second(s)
Bands
Specific: 47 kDa Non-specific: 44 kDa
Positive control
HEK293 cells transfected with S1P1 receptors were used (Lane 4). A clear band is found at ~47 kDa but it is above the bands found in mouse spinal cord, i.e. in Lanes 2 and 3. It is uncertain whether this band is S1P1.
Additional data
Abcam response
Thank you for your valuable feedback. We are currently investigating this negative review, and we will resolve this issue in accordance with our Abpromise guarantee.
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
投稿 Sep 21 2011