Application
Western blot
Sample
Human Cell lysate - whole cell (Vascular Smooth Muscle)
Gel Running Conditions
Reduced Denaturing (10% Bis-Tris with MOPS running buffer)
Loading amount
20 µg
Specification
Vascular Smooth Muscle
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Other product details
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 5% Milk
Dilution
1/2000
Secondary antibody
Dilution
1/10000
Name
Non-Abcam antibody was used: Donkey anti-goat IgG
Host species: Donkey
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Donkey
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Detection
Detection method
Femto
Exposure
2 minute(s) and 0 second(s)
Bands
Specific: 26 kDa
Additional data
Additional Notes
Comes up a little larger than expected (26kDa vs 21kDa), but this is quoted in the datasheet. A bit of a dirty Ab (ie high background), but doubling washing time after primary incubation from 30min to 60min gave a much cleaner blot. These are 4 different samples loaded evenly according to a BCA assay and then even loading confirmed by GAPDH loading control.
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
提交于 Jul 24 2008