Application
Western blot
Sample
Rat Cell lysate - whole cell (B50 Cell line)
Loading amount
6 µg
Specification
B50 Cell line
Gel Running Conditions
Non-reduced Denaturing (10%)
Blocking step
Milk as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Other product details
Dilution
1/1000
Incubation time
1 hour(s) and 0 minute(s) · Temperature: 24°C · Diluent: 5% Milk
Secondary antibody
Name
Non-Abcam antibody was used: Stratech Donkey Anti Rabbit HRP
Host species: Donkey
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Donkey
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/1000
Detection
Detection method
ECL
Exposure
10 minute(s) and 0 second(s)
Bands
Specific: 200 kDa Non-specific: 250 kDa
Positive control
This cell line is known to express Neurofilament Heavy
Negative control
Without primary (not shown)
Additional data
Additional Notes
Whole cell lysate was prepared in lysis buffer (Abcam) containing protease inhibitors.
6ug of lysate non-reduced (NR) run on 10% gel, then transferred to nitrocellulose membrane.
Block in 5% milk overnight at 4deg.
Incubate with primary for 1 hour at RT
Wash 3x10mins PBST
Incubate with secondary for 1hour at RT
Wash 3x10mins PBST
Develop using ECL.
6ug of lysate non-reduced (NR) run on 10% gel, then transferred to nitrocellulose membrane.
Block in 5% milk overnight at 4deg.
Incubate with primary for 1 hour at RT
Wash 3x10mins PBST
Incubate with secondary for 1hour at RT
Wash 3x10mins PBST
Develop using ECL.
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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提交于 Jul 12 2012