Western blot analysis of extracts from various cell lines using OMA1 (D4J7K) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). Mature OMA1 is smaller than its precursor, which has a calculated molecular weight of 59 kDa.Learn more about how we get our images.
For western blots, incubate membrane with diluted primary antibody in 5% w/v nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
NOTE: Please refer to primary antibody datasheet or product webpage for recommended antibody dilution.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).
NOTE: Volumes are for 10 cm x 10 cm (100 cm2) of membrane; for different sized membranes, adjust volumes accordingly.
* Avoid repeated exposure to skin.
posted June 2005
revised November 2013
Reprobing of an existing membrane is a convenient means to immunoblot for multiple proteins independently when only a limited amount of sample is available. It should be noted that for the best possible results a fresh blot is always recommended. Reprobing can be a valuable method but with each reprobing of a blot there is potential for increased background signal. Additionally, it is recommended that you verify the removal of the first antibody complex prior to reprobing so that signal attributed to binding of the new antibody is not leftover signal from the first immunoblotting experiment. This can be done by re-exposing the blot to ECL reagents and making sure there is no signal prior to adding the next primary antibody.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalently purified water.
posted June 2005
revised June 2016
Protocol Id: 263
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.
OMA1 (D4J7K) Rabbit mAb recognizes endogenous levels of total OMA1 protein.
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu504 of human OMA1 protein.
Mitochondria continuously divide and fuse. This dynamic process is highly regulated in response to various physiological cues (1,2). The GTPase OPA1 mediates the fusion of the mitochondrial inner membrane. Constitutive proteolytic processes mediated by OMA1 (S1 site) and YME1L (S2 site) convert long isoforms (L-OPA1) into short isforms (S-OPA1). The balance between L-OPA1 and S-OPA1 is required to maintain a normal morphology of mitochondria (3,4).
OMA1 is synthesized as a precursor and processed into a mature form (5,6). OMA1 is constitutively active and cleaves L-OPA1 at the S1 site. However, various stress stimuli can further activate OMA1 and result in the rapid and complete conversion of L-OPA1 into S-OPA1, which inhibits fusion and causes mitochondrial fragmentation (7).
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