Application
Western blot
Sample
Mouse Cell lysate - whole cell (cervix)
Loading amount
30 µg
Specification
cervix
Gel Running Conditions
Non-reduced Denaturing (10%)
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Other product details
Dilution
1/100
Incubation time
8 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 5% milk/TBST
Secondary antibody
Name
Non-Abcam antibody was used: Jackson Immuno Research Laboratories
Host species: Mouse
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Mouse
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/5000
Detection
Detection method
Super Signal
Exposure
20 minute(s) and 0 second(s)
Positive control
GAPDH
Additional data
Additional Notes
Tried to detect endogenous and overexpressed GAS 41 and no signal was observed. GAPDH bands were clear and present. Also tried 1:50, 1:500, and 1:1000 dilutions for endogenous and 1:500 for overexpression still with no bands. ECL was also tried with up to a 20 minute exposure before using supersignal. In the image, Lane 1 is over expression of GFP and Lane 2 is over expression of GAS 41. GFP was used to check transfection efficiency.
Abcam response
Thank you for your review of this untested species. You may be able to improve the signal by using more lysate or the nuclear fraction rather than whole cell lysate.
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.
DR. Angelique Whitehurst
Verified customer
投稿 Aug 17 2011