Western blot analysis of HeLa and THP-1 cell extracts, untreated (-) or treated with cycloheximide (Chx) #2112 (10 μg/mL, 16 hr) followed by human TNFα (TNFα) #8902 (20 ng/mL, 4hr) using #9496 Cleaved Caspase-8 (Asp391) (18C8) Rabbit mAb (left), or #9746 Caspase-8 (1C12) Mouse mAb (right). A band of unknown origin is detected at 25 kDa in some cell lines.
|MW (kDa)||18, 41, 43|
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
Cleaved Caspase-8 (Asp391) (18C8) Rabbit mAb detects human Caspase-8 when cleaved at Asp391 (isoform A, Asp374 on isoform B). This antibody will detect cleavage products containing the pro-domain with the p18 subunit as well as the p18 subunit alone. A band of unknown origin is detected at 25 kDa in some cell lines.
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues adjacent to Asp391 of human Caspase-8.
Apoptosis induced through the CD95 receptor (Fas/APO-1) and tumor necrosis factor receptor 1 (TNFR1) activates caspase-8 and leads to the release of the caspase-8 active fragments, p18 and p10 (1-3). Activated caspase-8 cleaves and activates downstream effector caspases such as caspase-1, -3, -6, and -7. Caspase-3 ultimately elicits the morphological hallmarks of apoptosis, including DNA fragmentation and cell shrinkage.
Explore pathways + proteins related to this product.