Anti-Caspase-8 Polyclonal Antibody is a Rabbit antibody targeting Caspase-8. Anti-Caspase-8 Polyclonal Antibody can be used in FCM,IF,IHC-Fr,IHC-P,WB.

IgG

Human,Rat (predicted:Mouse,Dog,Pig,Cow,Horse)

1. Blank control (blue line): U251 (blue).
Primary Antibody (green line): Rabbit Anti-caspase-8 antibody (TMAB-00290)
Dilution: 1 μg/10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody (white blue line): Goat anti-rabbit IgG-PE
Dilution: 1 μg/test.
Protocol
The cells were fixed with 70% ethanol overnight at 4°C and then permeabilized with 0.1% PBS-Tween for 20 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The cells were then incubated in 1 X PBS/2% BSA/10% goat serum to block non-specific protein-protein interactions followed by the antibody for 15 min at room temperature. The secondary antibody used for 40 min at room temperature.
2. Paraformaldehyde-fixed, paraffin embedded (rat liver); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (Caspase 8) Polyclonal Antibody, Unconjugated (TMAB-00290) at 1:500 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
3. Sample:
Jurkat (Human) Cell Lysate at 30 μg
K562 (Human) Cell Lysate at 30 μg
U87Mg (Human) Cell Lysate at 30 μg
Primary: Anti-caspase-8 subunit p18 (TMAB-00290) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 55/18 kDa
Observed band size: 57 kDa

WB: 1:500-2000; IHC-P: 1:100-500; IHC-Fr: 1:100-500; IF: 1:50-200; FCM: 1μg /Test

Polyclonal

KLH conjugated synthetic peptide: rat Caspase-8 subunit p10

Rat

CASP8

Caspase-8

Caspases,Metabolism,Necroptosis,Caspases,Caspases,Other proteases,Apoptosis

Most upstream protease of the activation cascade of caspases responsible for the TNFRSF6/FAS mediated and TNFRSF1A induced cell death. Binding to the adapter molecule FADD recruits it to either receptor. The resulting aggregate called death-inducing signaling complex (DISC) performs CASP8 proteolytic activation. The active dimeric enzyme is then liberated from the DISC and free to activate downstream apoptotic proteases. Proteolytic fragments of the N-terminal propeptide (termed CAP3, CAP5 and CAP6) are likely retained in the DISC. Cleaves and activates CASP3, CASP4, CASP6, CASP7, CASP9 and CASP10. May participate in the GZMB apoptotic pathways. Cleaves ADPRT. Hydrolyzes the small-molecule substrate, Ac-Asp-Glu-Val-Asp-|-AMC. Likely target for the cowpox virus CRMA death inhibitory protein. Isoform 5, isoform 6, isoform 7 and isoform 8 lack the catalytic site and may interfere with the pro-apoptotic activity of the complex.