Application
Western blot
Sample
Human Cell lysate - whole cell (Dopaminergic Neurons, HEK293T and HeLa)
Gel Running Conditions
Reduced Denaturing (NuPage 4-12% MES buffer)
Loading amount
30 µg
Specification
Dopaminergic Neurons, HEK293T and HeLa
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Other product details
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 5% Milk TBST
Dilution
1/1000
Secondary antibody
Dilution
1/20000
Name
Non-Abcam antibody was used: IRDye® 800CW Goat anti-mouse IgG
Host species: Goat
Clonality: Polyclonal
Conjugation: IRDye® 800CW
Host species: Goat
Clonality: Polyclonal
Conjugation: IRDye® 800CW
Detection
Detection method
LI-COR Odyssey CLx
Exposure
5 minute(s) and 0 second(s)
Bands
Specific: - kDa Non-specific: 48 kDa
Additional data
Additional Notes
human Embryonic Stem Cell-derived - 90 days differentiation.
Might not be strongly expressed in dopaminergic neurons. No signal detected in HEK293T or HeLa cells in the 90 kDa range. Might not be suitable for immunoblotting.
Abcam response
We would like to thank the reviewer for submitting this AbReview. We are sorry to hear that this antibody has given you inconclusive results in WB using differentiated human dopaminergic neurons, Hela cells and HEK293T cells. Based on our internal testing this antibody does not work in WB and as such has not been approved for use in Western Blotting. The submitted data is thus in line with our expectations.
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 Nov 20 2020