Western blot analysis of extracts from L-929 cells, untreated (-) or treated with Brefeldin A #9972 (10 μM, 30 hr; +), using Caspase-12 Antibody (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
|MW (kDa)||42, 55|
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not aliquot the antibody.
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
Caspase-12 Antibody recognizes endogenous levels of total full-length caspase-12 protein (55 kDa) and its cleaved product (42 kDa). Bands of unknown origin were detected at 12, 25, and 70 kDa.
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Lys163 of mouse caspase-12 protein. Antibodies are purified by protein A and peptide affinity chromatography.
Caspase-12 is located in the endoplasmic reticulum (ER). It is responsible for ER stress-induced apoptosis, such as high calcium concentration, low oxygen, and low glucose levels (1,2). One of the mechanisms for caspase-12 activation is related to calpain-mediated cleavage at T132 and K158, both of which are located at the amino-terminal region of caspase-12 (2,3). Caspase-12 also has a putative caspase cleavage site located at the carboxy-terminal region of the protein (3). In cortical neurons, caspase-12 is involved at least partially in the amyloid-beta neurotoxicity process (1).
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