Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing (15%)
Sample
Mouse Cell lysate - whole cell (MC3T3-E1 osteoblasts)
Specification
MC3T3-E1 osteoblasts
Treatment
Pre and post differentiation with osteogenic medium
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°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: Jackson immunoresearch
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/10000
Detection
Detection method
ECL
Exposure
1 minute(s) and 0 second(s)
Positive control
In the same samples, Histone H3 was stained using the same secondary antibody, and showed clear strong bands. These cells are known to express high levels of IGF-1.
Additional data
Additional Notes
Longer exposure had no effect - no bands were visible regardless. I also tried increasing sample concentration, and varying transfer times.
Ponceau staining showed protein is present on membrane.
Abcam response
Thank you for your feedback. Your results suggest this product is not suitable for western blot.
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. Benny Garfinkel
Verified customer
投稿 Aug 12 2013