Application
Western blot
Sample
Mouse Cell lysate - whole cell (BV2 cell line)
Gel Running Conditions
Reduced Denaturing (10% SDS-PAGE)
Loading amount
400000 cells
Treatment
LPS 50 ng/mL or LPS 50 ng/mL + UO 10 uM
Specification
BV2 cell line
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Other product details
Incubation time
18 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 5% Milk/PBS/0.05% Tween 20
Dilution
1/250
Secondary antibody
Dilution
1/3000
Name
Non-Abcam antibody was used: Santa Cruz Donkey anti-Goat-HRP
Host species: Donkey
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Donkey
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Detection
Detection method
ECL
Negative control
Medium treated cells
Exposure
5 minute(s) and 0 second(s)
Bands
Specific: 40-43 kDa kDa
Positive control
LPS treated cells
Additional data
Additional Notes
I would probably dilute this out more to 1:1000 and incubate for 48 hr. The 43kD band is strong and the other non-specific bands could disappear at this dilution for a clean band. Works better than another company's antibody!!
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 Jun 10 2008