Application
Western blot
Sample
Human Cell lysate - other (osteosarcoma)
Gel Running Conditions
Reduced Denaturing (12)
Loading amount
50 µg
Specification
osteosarcoma
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Other product details
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: TBS 1X Tween 0.05% milk 5%
Dilution
1/1000
Secondary antibody
Dilution
1/5000
Name
Non-Abcam antibody was used: anti-mouse
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Detection
Detection method
ECL+
Negative control
NDUFS3 KO cells
Exposure
1 minute(s) and 0 second(s)
Bands
Specific: 30 kDa Non-specific: 40, 22 kDa
Positive control
NDUFS3 WT cells
Additional data
Additional Notes
On nitrocellulose membrane the antibody recognizes the specific NDUFS3 band (verified by knocking out NDUFS3 by genome editing) but also two aspecific bands evident both in KO and wild type cellson whole cell lysates. The specific band only is detected in mitochondria. Samples prepared with proteases and phosphatases inhibitors.
Sample loading:
1. whole cell lysates from NDUFS3 KO cells
2. whole cell lysates from NDUFS3 KO cells
3. whole cell lysates from NDUFS3 WT cells
4. whole cell lysates from NDUFS3 WT cells
5. isolated mitochondria from NDUFS3 KO cells
6. isolated mitochondria from NDUFS3 KO cells
7. isolated mitochondria from NDUFS3 WT cells
8. isolated mitochondria from NDUFS3 WT cells
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 Nov 03 2015