Application
Western blot
Sample
Human Cell lysate - whole cell (kidney, epithelial cells)
Loading amount
100000 cells
Specification
kidney, epithelial cells
Treatment
transfected (or not) with GFP-M1 or pcDNA-NP or a minireplicon system and infected (or not) with influenza A virus
Gel Running Conditions
Reduced Denaturing (15%)
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 4% · Temperature: 18°C
Other product details
Dilution
1/500
Incubation time
1 hour(s) and 0 minute(s) · Temperature: 18°C · Diluent: PBS/tween
Secondary antibody
Name
Non-Abcam antibody was used: Li-Cor donkey anti-goat
Host species: Donkey
Clonality: Polyclonal
Conjugation: IRDye® 800CW
Host species: Donkey
Clonality: Polyclonal
Conjugation: IRDye® 800CW
Dilution
1/10000
Detection
Detection method
IR LI-cor
Exposure
2 second(s)
Bands
Specific: 27 and 54 kDa
Positive control
lane 1 and 2, cells transfected with GFP-M1, either infected (lane 1) or mock infected (lane 2).
Lane 5: Infected cells
Lane 7: M1 expressed from a minireplicon context
Lane 5: Infected cells
Lane 7: M1 expressed from a minireplicon context
Negative control
mock infected cells: lanes 4 and 6
lane 3 and 4: cells transfected with pcDNA-NP
lanes 6: mock infected cells
lane 8: M1 expressed from a minireplicon system that lacks a viral polymerase.
lane 3 and 4: cells transfected with pcDNA-NP
lanes 6: mock infected cells
lane 8: M1 expressed from a minireplicon system that lacks a viral polymerase.
Additional data
Additional Notes
This antibody detected M1 (and GFP-M1), either from infected cells and in the context of a minireplicon system. It did not detect NP, and lanes with mock infected samples were clean.
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.
DR. Maria Amorim
Verified customer
提交于 Mar 27 2012