Application
Western blot
Sample
Mouse Cell lysate - whole cell (mouse embryonic fibroblasts)
Gel Running Conditions
Reduced Denaturing
Loading amount
30 µg
Specification
mouse embryonic fibroblasts
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Other product details
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C
Dilution
1/500
Secondary antibody
Name
Non-Abcam antibody was used: goat ant-rabbit sc2004 from Sanatcruz
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Detection
Detection method
ECL
Exposure
4 minute(s) and 0 second(s)
Bands
Specific: 25 kDa
Additional data
Additional Notes
Although the band closet to 33 kDA which was very clear it was at 25kDa instead of 33kDa.
Abcam response
Thank you for sharing your feedback with us. There is some varaibility in a Western Blot, and you may sometimes see a protein migrate up to 10 kD from the predicted based on post-translational modifications, splice variants, and charge.
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 Jul 29 2014