Application
Western blot
Sample
Human Cell lysate - whole cell (human astrocyte)
Loading amount
25 µg
Specification
human astrocyte
Treatment
treated with LXR ligand for 20 hrs
Gel Running Conditions
Reduced Denaturing (4-15% gradient gel)
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Other product details
Dilution
1/1500
Incubation time
17 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 1% milk in TBST
Secondary antibody
Name
Non-Abcam antibody was used: Bio-Rad IgG (H+L) HRP conjugate
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/5000
Detection
Detection method
Supersignal West Pico
Exposure
30 second(s)
Bands
Specific: 240 kDa Non-specific: 50, 90, 110, 300 kDa
Additional data
Additional Notes
The samples should not be heated. Instead after the addition of sample buffer, those were left on the RT for O/N. Cell lysis buffer contains 1% TX100. Based on the intensity of specific band, even higher dilution may be used.
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 Aug 06 2009