Western blot analysis of extracts from various cell lines using Caspase-14 (D3J3D) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing full-length human caspase-14 protein (hCaspase-14; +), using Caspase-14 (D3J3D) Rabbit mAb.
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.
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-14 (D3J3D) Rabbit mAb recognizes endogenous levels of total caspase-14 protein. Endogenous levels of the p20 activated subunit have not been detected.
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ala71 of human caspase-14 protein.
Caspases are a family of cysteine proteases that play an essential role in carrying out apoptosis. Caspase-14, also named MICE, is a unique member of the caspase family with restricted expression; it is found in embryonic tissues and adult skin (1,2). Caspase-14 is weakly processed into p18 and p11 subunits by caspase-8 (2). Caspase-14 may not play a role in apoptosis (2), but instead may regulate keratinocyte differentiation (3). Expression of caspase-14 may protect from psoriasis (4,5) and irradiation damage (6). Caspase-14 may also be responsible for proteolytic processing of filaggrin during terminal differentiation of keratinocytes (6).
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