Application
Western blot
Sample
Mouse Cell lysate - whole cell (RAW 264.7 murine macrophage cell line)
Loading amount
50 µg
Specification
RAW 264.7 murine macrophage cell line
Treatment
IL-4 10 ng.mL-1 for 48 hours
Gel Running Conditions
Reduced Denaturing (10% acrylamyde gel)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Other product details
Dilution
1/1000
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 3% milk in PBS
Secondary antibody
Name
Non-Abcam antibody was used: ImmunoPure Rabbit Anti-Goat IgG, (H+L),
Host species: Rabbit
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Rabbit
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/10000
Detection
Detection method
ECL Millipore
Exposure
10 second(s)
Bands
Specific: ˜37 kDa Non-specific: ˜30 kDa
Positive control
Mouse liver whole cell extract (lane 1)
Negative control
RAW cells untreated (lane 2)
Additional data
Additional Notes
PVDF membrane activated with methanol, 1½ hour, 100 V, 4°C, RIPA lysis buffer.
Lanes:
1. Mouse liver extract
2. Untreated RAW cells
3. IL-4 treated RAW cells
Lanes:
1. Mouse liver extract
2. Untreated RAW cells
3. IL-4 treated RAW cells
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投稿 Jan 22 2010