Application
Western blot
Sample
Mouse Cell lysate - whole cell (Mouse (macrophage, skeletal muscle) Human (kidney))
Gel Running Conditions
Reduced Denaturing (8-18% gradient)
Loading amount
5 µg
Treatment
293A cells were treated with mTORC1 catalytic inhibitor 50nM for 1hr to inhibit S6 phosphorylation
Specification
Mouse (macrophage, skeletal muscle) Human (kidney)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Other product details
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: TBS-T
Dilution
1/1000
Secondary antibody
Dilution
1/10000
Name
Non-Abcam antibody was used: Goat anti-rabbbit whole IgG secondary
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Detection
Detection method
ECL+
Negative control
293A cells with mTORC1 inhibition to reduce pS6 phosphorylation.
Exposure
5 minute(s) and 0 second(s)
Bands
Specific: 32 kDa
Positive control
293A cells, mouse bone marrow derived macrophages (BMDM).
We were looking for an antibody that works in skeletal muscle. Unfortunately, non-of the commercial antibodies work well, leading us to believe this signalling is not robust in murine muscle, likely not an antibody problem.
Additional data
Abcam response
Thank you for your valuable feedback.
This antibody has not previously been tested in mouse and we are pleased to see this has been successful. The alignment of the mouse protein with the immunogen is over 90%.
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 Jun 21 2017