For SDS PAGE experiments, 20μg of mitochondrial proteins or 15μg of whole cell extracts were loaded in 15% SDS gels. Membranes were probed with the following antibodies: NDUFA9 (1:1000, abcam, ab14713), NDUFAF1 (1:10000, abcam, ab79826), NDUFS3 (1:1000, abcam, ab110246), NDUFB9 (1:1000, abcam, ab106699), NDUFV2 (1:1000, proteintech, 15301-1-AP), Prohibitin (1:1000, abcam, ab28172), SDHA (1:1000, abcam, ab14715), UQCRC2 (1:1000, abcam, ab14745), Tubulin (1:1000, Cell Signaling, #21485). Proteins separated by BN PAGE were transferred to PVDF membranes and western blotting was performed as above. Alternatively, the BN gels were used for in-gel activity assays for complex I by incubating the gel in the assay buffer consisting of 5mM Tris/HCl with 2.5mg/ml nitrotetrazolium blue and 0.1mg/ml NADH (pH 7.4).
Ab14745
Manufactured by Cell Signaling Technology
Ab14745 is a recombinant monoclonal antibody that recognizes the human ATRX protein. This antibody is designed for use in Western blotting, immunoprecipitation, and immunohistochemistry applications.
Automatically generated - may contain errors
2 protocols using ab14745
1
Mitochondrial Protein Analysis by SDS-PAGE and BN-PAGE
2
Mitochondrial Protein Analysis by SDS-PAGE and BN-PAGE
Check if the same lab product or an alternative is used in the 5 most similar protocols
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For SDS–PAGE experiments, 20 μg of mitochondrial
proteins or 15 μg of whole-cell extracts were loaded in 15%
SDS gels. Membranes were probed with the following antibodies: NDUFA9 (1:1,000, Abcam, ab14713),
NDUFAF1 (1:10,000, Abcam,
ab79826), NDUFS3 (1:1,000,
Abcam, ab110246), NDUFB9
(1:1,000, Abcam, ab106699), NDUFV2 (1:1,000, Proteintech, 15301-1-AP), PhB (1:1,000, Abcam, ab28172),
SDHA (1:1,000, Abcam,
ab14715), UQCRC2 (1:1,000,
Abcam, ab14745) and tubulin (1:1,000, Cell Signaling, no. 21485). Proteins
separated by BN PAGE were transferred to Polyvinylidene fluoride (PVDF)
membranes and western blotting was performed as above. Alternatively, the BN
gels were used for in-gel activity assays for complex I by incubating the gel in
the assay buffer consisting of 5 mM Tris/HCl with
2.5 mg ml−1nitrotetrazolium blue and
0.1 mg ml−1NADH (pH 7.4).
proteins or 15 μg of whole-cell extracts were loaded in 15%
SDS gels. Membranes were probed with the following antibodies: NDUFA9 (1:1,000, Abcam, ab14713),
NDUFAF1 (1:10,000, Abcam,
ab79826), NDUFS3 (1:1,000,
Abcam, ab110246), NDUFB9
(1:1,000, Abcam, ab106699), NDUFV2 (1:1,000, Proteintech, 15301-1-AP), PhB (1:1,000, Abcam, ab28172),
SDHA (1:1,000, Abcam,
ab14715), UQCRC2 (1:1,000,
Abcam, ab14745) and tubulin (1:1,000, Cell Signaling, no. 21485). Proteins
separated by BN PAGE were transferred to Polyvinylidene fluoride (PVDF)
membranes and western blotting was performed as above. Alternatively, the BN
gels were used for in-gel activity assays for complex I by incubating the gel in
the assay buffer consisting of 5 mM Tris/HCl with
2.5 mg ml−1nitrotetrazolium blue and
0.1 mg ml−1NADH (pH 7.4).
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