WB result of Caspase-8 Recombinant Rabbit mAb
Primary antibody: Caspase-8 Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: HCT 116 whole cell lysate 20 µg
Lane 2: HeLa whole cell lysate 20 µg
Lane 3: THP-1 whole cell lysate 20 µg
Lane 4: Jurkat whole cell lysate 20 µg
Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution Predicted MW: 55 kDa
Observed MW: 58 kDa
Caspase-8 Recombinant Rabbit mAb (S-1540-1)
Caspase-8 Recombinant Rabbit mAb (S-1540-1)
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Product Details
Product Details
Product Specification
| Host | Rabbit |
| Antigen | Caspase-8 |
| Synonyms | CASP-8; Apoptotic cysteine protease; Apoptotic protease Mch-5; CAP4; FADD-homologous ICE/ced-3-like protease; FADD-like ICE (FLICE); ICE-like apoptotic protease 5; MORT1-associated ced-3 homolog (MACH); CASP8; MCH5 |
| Immunogen | Synthetic Peptide |
| Location | Cytoplasm, Nucleus, Cell projection |
| Accession | Q14790 |
| Clone Number | S-1540-1 |
| Antibody Type | Recombinant mAb |
| Isotype | IgG |
| Application | WB, ICC, ICFCM |
| Reactivity | Hu |
| Positive Sample | HCT 116, HeLa, THP-1, Jurkat |
| Purification | Protein A |
| Concentration | 0.5 mg/ml |
| Conjugation | Unconjugated |
| Physical Appearance | Liquid |
| Storage Buffer | PBS, 40% Glycerol, 0.05% BSA, 0.03% Proclin 300 |
| Stability & Storage | 12 months from date of receipt / reconstitution, -20 °C as supplied |
Dilution
| application | dilution | species |
| WB | 1:1000 | Hu |
| ICC | 1:500 | Hu |
| ICFCM | 1:50 | Hu |
Background
Caspase-8 is a member of the cysteine-aspartic acid protease (caspase) family. Sequential activation of caspases plays a central role in the execution-phase of cell apoptosis. Caspases exist as inactive proenzymes composed of a prodomain, a large protease subunit, and a small protease subunit. Activation of caspases requires proteolytic processing at conserved internal aspartic residues to generate a heterodimeric enzyme consisting of the large and small subunits. This protein is involved in the programmed cell death induced by Fas and various apoptotic stimuli. The N-terminal FADD-like death effector domain of this protein suggests that it may interact with Fas-interacting protein FADD. This protein was detected in the insoluble fraction of the affected brain region from Huntington disease patients but not in those from normal controls, which implicated the role in neurodegenerative diseases.
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Western Blot
FC
Flow cytometric analysis of 4% PFA fixed 90% methanol permeabilized Jurkat (Human T cell leukemia T lymphocyte) labelling Caspase-8 antibody at 1/50 dilution (1 μg) / (Red) compared with a Rabbit monoclonal IgG (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti - Rabbit IgG Alexa Fluor® 488 was used as the secondary antibody.
Immunocytochemistry
ICC shows positive staining in Jurkat cells. Anti- Caspase-8 antibody was used at 1/500 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 4% PFA and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue). Counterstain with tubulin (Red).
