|MW (kDa)||46, 43, 28|
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: 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 with the heavy chain subunit of human Cathepsin D. Antibodies are purified by protein A and peptide affinity chromatography.
Cathepsin D is a ubiquitously expressed lysosomal aspartyl protease involved in the normal degradation of proteins (1). It is synthesized as an inactive 43 kDa preprocathepsin D that is cleaved and glycosylated to form a 46 kDa procathepsin D and then further cleaved to produce 28 kDa and 15 kDa subunits (heavy and light chains, respectively) (2). Cathepsin D may also be secreted into the cytosol during apoptosis and contribute to cleavage of substrates implicated in the apoptotic pathway (3). Numerous studies have suggested that cathepsin D plays a role in neuronal degradation and malignant transformation, particularly in breast cancer (4-9).
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