Application
Western blot
Sample
Human Cell lysate - whole cell (Patient fibroblasts)
Gel Running Conditions
Reduced Denaturing (6 %)
Loading amount
15 µg
Specification
Patient 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
12 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: TBS + Tween
Dilution
1/1000
Secondary antibody
Dilution
1/5000
Name
Non-Abcam antibody was used: Amersham ECL Rabbit IgG, HRP-linked whole Ab (from
Host species: Donkey
Clonality: Monoclonal
Conjugation: HRP polymer
Host species: Donkey
Clonality: Monoclonal
Conjugation: HRP polymer
Detection
Detection method
Chemiluminescent
Exposure
2 minute(s) and 0 second(s)
Bands
Specific: ~500 kDa Non-specific: 250, 350 kDa
Additional data
Additional Notes
Do not heat the whole cell extracts.
Protein extraction was done with RIPA buffer + PI. After centrifugation (20 000 g/5min), the protein concentration was measured in supernatant.
Proteins were mixt with SDS sample buffer + DTT then loaded in 6 % SDS tris-glycine Gels
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.
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投稿 Jan 30 2017