Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing (4-12% gel)
Sample
Rat Tissue lysate - nuclear (Brain tissue homogenate, P1 and P2 fractions)
Specification
Brain tissue homogenate, P1 and P2 fractions
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Other product details
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 5% milk in PBS with 0.1% Tween-20
Dilution
1/1000
Secondary antibody
Name
Non-Abcam antibody was used: Goat anti-rabbit HRP (Sigma A6154)
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/1000
Detection
Detection method
ECL
Bands
Specific: 60 kDa Non-specific: 35 kDa
Exposure
5 minute(s) and 0 second(s)
Negative control
Fraction of tissue lysate minus nucleus
Positive control
Nuclear fraction of tissue lysate
Additional data
Additional Notes
Lane 1 = Crude homogenate 20 ´g
Lane 2 = Crude homogenate 40 ´g
Lane 3 = P1 (nuclear fraction) 20 ´g
Lane 4 = P1 (nuclear fraction) 40 ´g
Lane 5 = P2 (non-nuclear fraction) 20 ´g
Lane 6 = P2 (non-nuclear fraction) 40 ´g
Non-specific band at ~35 KDa only evident in non-nuclear fractions
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 Jan 31 2014