Application
Western blot
Loading amount
25 µg
Gel Running Conditions
Reduced Denaturing (12%)
Sample
African Green Monkey Cell lysate - whole cell (COS cells, HeLa cells (human), N2A cells (mouse),)
Specification
COS cells, HeLa cells (human), N2A cells (mouse),
Blocking step
LiCor blocking reagent as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 23°C
Other product details
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: LiCor blocking reagent
Dilution
1/1000
Secondary antibody
Name
Non-Abcam antibody was used: Li-Cor IRDye 680
Host species: Donkey
Clonality: Polyclonal
Conjugation: IRDye 680
Host species: Donkey
Clonality: Polyclonal
Conjugation: IRDye 680
Dilution
1/15000
Detection
Detection method
Odyssey Clx
Bands
Specific: 51 kDa
Exposure
6 minute(s) and 0 second(s)
Positive control
HeLa cell lysate - but did not observe bands
Additional data
Additional Notes
Image notes:
25ug per lane
1 – COS cell lysate (green monkey)
2 – COS cell lysate (green monkey)
3 – HeLa cell lysate (human)
4 – Neuro2a cell lysate (mouse)
5- Neuro2a cell lysate (mouse)
6 – mouse embryonic fibroblasts
7 – mouse embryonic fibroblasts
8 – COS cell mitochondrial fraction
9 – COS cell mitochondrial fraction
Only MEFs lanes 6 and 7 had correct band. Slightly smaller band in 8 and 9. Should have worked in HeLa cells but didn't see a band. May work in mouse.
Immobilon PVDF membrane and LiCor reagents and protocol used for WB.
Samples were lysed in RIPA buffer.
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
投稿 Dec 22 2014