Application
Western blot
Sample
Drosophila melanogaster Cell lysate - other (whole fly mitochondrial fraction)
Gel Running Conditions
Reduced Denaturing (10)
Loading amount
50 µg
Specification
whole fly 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
12 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: PBS 1% BSA
Concentration
0.5 µg/ml
Secondary antibody
Dilution
1/1000
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
whole fly cytoplasmic fraction
Exposure
1 minute(s) and 30 second(s)
Additional data
Additional Notes
This antibody works well with mouse and avian samples in both reduced and native conditions. However, the epitope this antibody binds appears to not be conserved in arthropods. I tried multiple samples on two different gels and tried incubating the primary for nearly two hours at room temperature or overnight at 4C. Under no conditions did I observe bands in the fly samples.
Mitochondrial and cytoplasmic fractions were obtained from whole flies and whole wolf spider with Abcam Mitochondrial Isolation Kit for Tissue (ab110168).
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 Sep 12 2017