Western blot analysis of extracts from KARPAS-299, THP-1 and IM-9 cells using Caspase-4 Antibody.
Western blot analyis of extracts from recombinant, active caspase-1, -4, and -5 using Caspase-4 Antibody.
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-4 Antibody detects endogenous levels of total caspase-4 protein. Processing intermediate forms of caspase-4 are observed at 40 kDa and 32 kDa as previously reported (7). The antibody does not cross-react with other caspases.
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ile125 within the p20 subunit of human caspase-4 protein. Antibodies were purified by protein A and peptide affinity chromatography.
Caspase-4 (TX/ICH-2/ICErelII) is a member of the caspase family of proteases that play a key role in the execution of apoptosis and activation of inflammatory cytokines (1-3). Expression of caspase-4 has been observed in most tissues except brain, with highest levels in placenta, lung, spleen, and peripheral blood lymphocytes (PBL). Caspase-4 was originally found to contribute to Fas-mediated apoptosis (4). Several caspases (including caspase-4, caspase-5, and mouse caspase-11 and -12) are most closely related to caspase-1 and are capable of inducing apoptosis when over-expressed but are better characterized in the proteolytic activation of inflammatory cytokines (5). Caspase-4 associates with TRAF6 and is involved in the LPS inducible production of inflammatory cytokines IL-8 and MIP1 in THP-1 cells (6). While caspase-4 and mouse caspase-12 localize to the endoplasmic reticulum (ER) and may be activated by drugs that induce ER-stress (7), at least one study suggests that caspase-4 and caspase-12 are not essential for the ER-stress induced apoptosis (8).
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