Application
Western blot
Sample
Drosophila melanogaster Tissue lysate - other (whole fly mitochondrial fraction)
Gel Running Conditions
Reduced Denaturing (18)
Loading amount
25 µg
Specification
whole fly mitochondrial fraction
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Other product details
Incubation time
1 hour(s) and 45 minute(s) · Temperature: 23°C · Diluent: PBS 1% BSA
Concentration
1 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
whole fly cytoplasmic fraction
Exposure
10 second(s)
Positive control
mitochondrial fraction from AML12 cells
Additional data
Additional Notes
Lane 1: Thermo Scientific PageRuler Prestained Protein Ladder
Lanes 2-3: Fly mitochondrial fraction
Lane 4: Fly cytoplasmic fraction
Lane 5: Robin mitochondrial fraction
Lane 6: Robin cytoplasmic fraction
Lane 7: Mouse (AML12) mitochondrial fraction
Lane 8: Mouse (AML12) cytoplasmic fraction
Lane 9: Bio-Rad Precision Plus Protein WesternC Standards
Mitochondrial and cytoplasmic fractions were obtained from whole flies with Abcam Mitochondrial Isolation Kit for Tissue (ab110168) and from robin fibroblast and mouse hepatocyte cell lines with Abcam Mitochondrial Isolation Kit for Cells (ab110170).
This antibody produces a strong and specific signal with mouse samples run on native and reducing gels. However, the targeted epitope does not appear to be present in the bird or fly proteins.
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 Aug 21 2017