Application
Western blot
Sample
Human Cell lysate - whole cell (kidney)
Gel Running Conditions
Reduced Denaturing (4-12% bis-tris gel, MES buffer)
Loading amount
30 µg
Specification
kidney
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
Dilution
1/1000
Secondary antibody
Dilution
1/15000
Name
Non-Abcam antibody was used: IRDye® 680RD Goat anti-Mouse IgG (H + L)
Host species: Goat
Clonality: Polyclonal
Conjugation: IRDye® 680RD
Host species: Goat
Clonality: Polyclonal
Conjugation: IRDye® 680RD
Detection
Detection method
OdysseyCLX
Exposure
5 minute(s) and 0 second(s)
Positive control
There was no positive control.
Additional data
Additional Notes
I didn't stimulate the 293T cells, thus it is possible that 293T cells do not have basal level of IL8. 30ug/well is the maximum loading amount that I used. I haven't tried to gather the media for the secreted IL8, and it will be worth trying in the future.
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 Nov 03 2020