Application
Western blot
Sample
Mouse Tissue lysate - whole (lungs)
Gel Running Conditions
Reduced Denaturing (30 minutes RT, 10% gel)
Loading amount
100 µg
Specification
lungs
Blocking step
2 % Topblock in TBT-0.1% Triton 100 x as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
Other product details
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C
Dilution
1/1000
Secondary antibody
Dilution
1/5000
Name
Non-Abcam antibody was used: promega
Host species: Donkey
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Donkey
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Detection
Detection method
ECL
Negative control
used tissue from knockout nrf2 mice. Lane 7
Exposure
3 minute(s) and 0 second(s)
Bands
Specific: 100 (expected) kDa Non-specific: 68, 100180, 55, 35, 2 kDa
Positive control
used wild type mice with acute inflammation. Lane 6
Additional data
Additional Notes
Tissue was lysate wth RIPA buffer+PMSF+protease inhibitor cocktail. Also compared the ab76026 with ab62352. Could not detect specific bands with the conditions used.
lanes:
1and 5 Wild type healthy
2 and 6 wild type inflammation
3 and 7 nrf2 KO healthy
4 and 8 nrf2 KO inflammation
1-4 : ab62352
5-8: ab76026
Abcam response
We would like to thank this researcher for submitting their review. This antibody had not been tested for cross reactivity with mouse and we were unsure of what the results would be. The reviewer received an AbTrial discount code for using the antibody in an untested species.
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 Dec 13 2016