Western blot abreview for Anti-MLKL antibody [EPR17514]

Excellent
Abreviews
Application
Western blot
Sample
Human Cell lysate - whole cell (HeLa, HT29, L929, MEF)
Gel Running Conditions
Reduced Denaturing (10%)
Loading amount
10 µg
Specification
HeLa, HT29, L929, MEF
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Other product details

Incubation time
1 hour(s) and 0 minute(s) · Temperature: 22°C · Diluent: 5% BSA in TBST
Dilution
1/1000

Secondary antibody

Dilution
1/10000
Name
Non-Abcam antibody was used: Sigma antibody
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase

Detection

Detection method
ECL
Negative control
human: HeLa MLKL KO cell mouse: L929 MLKL KO cell
Exposure
30 second(s)
Bands
Specific: 54 kDa
Positive control
human: HeLa and HT29 WT cell mouse: L929 and MEF WT cell

Additional data

Additional Notes
This antibody works well in detection of human MLKL and mouse MLKL. But in L929 and MEF cell lysates, there is one nonspecific band slightly higher than mouse MLKL.
Abcam response
Thank you very much for your valuable feedback. We are pleased that the Anti-MLKL antibody ab184718 is working well for you in human and mouse samples. We are currently unsure about the nature of the unspecific signal slightly higher than MLKL in the mouse cell lines L929 and MEF. Further dilution of the antibody or blocking with milk may reduce the unspecific binding. We would like to encourage our customer to share their experience with the MLKL antibody in mouse samples.

Xiaozheng Xu

Verified customer

Submitted Sep 08 2015

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