Application
Western blot
Sample
Mouse Cell lysate - whole cell (Primary MEF)
Loading amount
5 µg
Specification
Primary MEF
Treatment
100ngµl TPA for 2 hours
Gel Running Conditions
Reduced Denaturing (4-14% Bis-Tris)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Other product details
Dilution
1/1000
Incubation time
14 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: PBST 0.1% + 4% milk
Secondary antibody
Name
Non-Abcam antibody was used: Anti-rabbit IgG (H&L) IRDye 800
Host species: Goat
Clonality: Polyclonal
Conjugation: IRDye® 800CW
Host species: Goat
Clonality: Polyclonal
Conjugation: IRDye® 800CW
Dilution
1/10000
Detection
Detection method
Fluorescence (LICOR Odyssey)
Exposure
1 minute(s) and 0 second(s)
Bands
Specific: none kDa Non-specific: 15, 25, 50, 59-80 kDa
Positive control
Wild type primary MEFs + 100ngµl TPA for 2 hours. As we predicted anti-DUSP7 may cross react with DUSP6 (MKP3) protein we also used MKP-3 null primary MEFs + 100ngµl TPA for 2 hours.
Negative control
Unstimulated wild type primary MEFs
Additional data
Abcam response
Thank you for sharing this data with us. We have not previously tested this antibody on mouse lysates and from this abreview it would appear that the antibody is not suitable for western blot on mouse samples, under these conditions. Abcam would welcome feedback from other customers using this 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.
Abcam user community
Verified customer
提交于 Aug 28 2012