Application
Western blot
Sample
Mouse Cell lysate - other (Fibroblasts, mitochondrial fraction.)
Gel Running Conditions
Reduced Denaturing (18)
Loading amount
25 µg
Specification
Fibroblasts, mitochondrial fraction.
Blocking step
BSA as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Other product details
Incubation time
1 hour(s) and 45 minute(s) · Temperature: 23°C · Diluent: PBS 1% BSA
Concentration
0.5 µg/ml
Secondary antibody
Dilution
1/3000
Name
Non-Abcam antibody was used: Cell Signaling Technology anti-Mouse IgG #7076
Host species: Horse
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Horse
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Detection
Detection method
SuperSignal Pico ECL
Negative control
Cytoplasmic fraction.
Exposure
10 second(s)
Bands
Specific: 35 kDa
Additional data
Additional Notes
The bands around 75 kDa are Anti-SDHA [2E3GC12FB2AE2] ab14715 which was not removed by the stripping buffer.
Mitochondrial and cytoplasmic fractions were obtained from robin fibroblast and mouse hepatocyte (AML12) cell lines with Abcam Mitochondrial Isolation Kit for Cells (ab110170).
This antibody works well on samples run on both reducing and native gels.
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 Sep 06 2017