Application
Western blot
Sample
Human Cell lysate - whole cell (Hela, HT29)
Loading amount
30 µg
Specification
Hela, HT29
Treatment
see legend
Gel Running Conditions
Reduced Denaturing (7% Tris acetate)
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Other product details
Dilution
1/20000
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 5% milk
Secondary antibody
Name
Non-Abcam antibody was used: goat anti rabbit HRP
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/10000
Detection
Detection method
ECL
Exposure
10 minute(s) and 0 second(s)
Bands
Specific: 65 kDa Non-specific: none kDa
Positive control
Hela cells with or without etoposide treatment. Etoposide induces CHk2 phosphorylation as can be seen with band shift in lane 4. This is why a little blurry as used 7% gel in order to see BS and show antibody recognises phospho form (not all out there do!!)
Negative control
Chk2 siRNA treated cells
Additional data
Additional Notes
Image legend:
1: HT29 cells mock treated
2: HT29 cells Chk2 siRNA
3: HeLa cells
4: HeLa cells plus 50ug/ml etoposide, 6 hours
5: HT29 cells mock
6: HT29 cells Chk2 siRNA
LC = Loading control. MSH2
ECL imaged on Li-Cor instrument
1: HT29 cells mock treated
2: HT29 cells Chk2 siRNA
3: HeLa cells
4: HeLa cells plus 50ug/ml etoposide, 6 hours
5: HT29 cells mock
6: HT29 cells Chk2 siRNA
LC = Loading control. MSH2
ECL imaged on Li-Cor instrument
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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提交于 May 02 2013