For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
NOTE: Please refer to primary antibody product webpage for recommended antibody dilution.
From sample preparation to detection, the reagents you need for your Western Blot are now in one convenient kit: #12957 Western Blotting Application Solutions Kit
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: Loading of prestained molecular weight markers (#59329, 10 µl/lane) to verify electrotransfer and biotinylated protein ladder (#7727, 10 µl/lane) to determine molecular weights are recommended.
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 June 2020
Protocol Id: 10
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human MT-ND3 protein. Antibodies are purified by peptide affinity chromatography.
Mitochondrially encoded NADH dehydrogenase 3 (MT-ND3) is encoded by the mitochondrial gene MT-ND3 and is a core subunit of the mitochondrial membrane respiratory chain NADH dehydrogenase (Complex I). Located in the mitochondrial inner membrane, Complex I is the largest of the five respiratory complexes and catalyzes electron transfer from NADH to ubiquinone. The mitochondrially encoded subunits of Complex I, including MT-ND3, are the most hydrophobic subunits and form the core of the transmembrane region (1,2). Pathogenic variants of the MT-ND3 gene are known to cause mitochondrial complex I deficiency (MT-C1D) and may lead to a wide range of clinical disorders, including Leigh syndrome, Leber hereditary optic neuropathy, and encephalopathy (3-5).
UniProt ID: P03897
Entrez-Gene Id: 4537
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