Western blot abreview for Anti-ERK1 + ERK2 antibody

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Abreviews
Application
Western blot
Sample
Rat Tissue lysate - whole (spinal cord homogenate)
Loading amount
20 µg
Specification
spinal cord homogenate
Treatment
sham or nerve transected
Gel Running Conditions
Reduced Denaturing (10% SDS-PAGE)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Other product details

Dilution
1/1000
Incubation time
18 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 5% Milk/PBS/0.05% Tween 20

Secondary antibody

Name
Non-Abcam antibody was used: Pierce Goat anti-Rabbit-HRP
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/3000

Detection

Detection method
ECL
Exposure
5 minute(s) and 0 second(s)
Bands
Specific: 42, 44 kD kDa
Positive control
Nerve transected
Negative control
Sham operated

Additional data

Additional Notes
The rat tissue homogenate used here was from animals that underwent either Sham surgery or L5 spinal nerve transection. The tissue was harvested seven days post surgery, sonicatedwith RIPA buffer, protein estimate made by Lowry and 20 ug loaded in each lane. A 10% SDS-PAGE gel was run for 1.5 hr at 100V and transfered to PVDF membrane for 1.5 hr at 274 mA.

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Submitted Jun 18 2008

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