Caspase 8 antibody (AA 411-482)
- Target See all Caspase 8 (CASP8) Antibodies
- Caspase 8 (CASP8)
- AA 411-482
- Human, Mouse, Rat, Monkey, Pig, Sheep
- This Caspase 8 antibody is un-conjugated
- Western Blotting (WB), ELISA, Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Flow Cytometry (FACS), Immunofluorescence (Cultured Cells) (IF (cc)), Immunofluorescence (Paraffin-embedded Sections) (IF (p)), Immunohistochemistry (Frozen Sections) (IHC (fro))
- Predicted Reactivity
- Purified by Protein A.
- KLH conjugated synthetic peptide derived from mouse Caspase-8 subunit p10
- Application Notes
- For Research Use only
Validation #029759 (Western Blotting)Validation ImagesFull Methods
- Alamo Laboratories Inc
- Lot Number
- Method validated
- Western Blotting
- Positive Control
- HeLa cells
- Negative Control
- c6/36 mosquito cells (non-reactive species)
- A strong band was observed at the correct molecular weight in the positive control sample. No major bands were observed in the negative sample.
- Primary Antibody
- Antigen: Caspase 8, Apoptosis-Related Cysteine Peptidase (CASP8) (1:200 dilution)
- Catalog number: ABIN724205
- Lot number: 131127
- Secondary Antibody
- Antigen: Goat Anti-Rabbit IgG (H + L)-HRP Conjugate (1:20,000 dilution)
- Lot number: L170-6515
- Full Protocol
- 1. Total protein extracts were boiled in 1X SDS Sample Buffer containing 1% SDS and 1.25% β-mercaptoethanol at 95°C for 5 min prior to loading.
- 2. 20 μg of boiled extracts were loaded and resolved on 8-16% SDS-polyacrylamide gel.
- 3. The Thermo Scientific - Spectra Multicolor Broad Range (Cat # 26634) were used as molecular mass markers.
- 4. Proteins were then transferred onto PVDF membrane by wet transfer and protein transfer was confirmed with Ponceau-S staining.
- 5. The PVDF membrane was incubated with 25 mL of blocking buffer [Tris Buffered Saline, pH 7.4 plus 0.1% TW20 (TBST)] containing 5% (W/V) BSA at room temperature for 1 h.
- 6. The membrane was rinsed with TBST once.
- 7. The membrane was immersed with the protein side up in the primary antibody solution (CASP8; 1:200) in TBST containing 5% (W/V) BSA and incubated for 16 h at 4°C.
- 8. The membrane was rinsed in TBST thrice for 5 min each.
- 9. The membrane was incubated in the HRP-conjugated secondary antibody solution (Goat anti-rabbit IgG-HRP; 1:20,000) in TBST containing 5% (W/V) BSA and incubated for 1 hour at room temperature (~26°C) with gentle agitation.
- 10. The membrane was rinsed thrice TBST thrice for 5 min each.
- 11. The membrane was rinsed in TBS twice for 30 s each.
- 12. Signals were detected with ECL-2 Substrate. The blot was scanned for 300 s.
- 13. The membrane was rinsed three times TBST.
- 14. Incubated in Acidic Glycine Stripping Buffer at room temperature with gentle agitation for 3 times, 10 min each.
- 15. The membrane was washed in TBST 2 times for 10 min each.
- 16. Repeated Steps 5-12 with the loading control antibody (anti-Actin; 1:6,000) and its matching secondary antibody (Goat anti-rabbit IgG-HRP; 1:20,000).
- Experimental Notes
- - Nothing to note.
- 1 μg/μL
- 0.01M TBS( pH 7.4) with 1 % BSA, 0.03 % Proclin300 and 50 % Glycerol.
- Precaution of Use
- This product contains ProClin: a POISONOUS AND HAZARDOUS SUBSTANCE, which should be handled by trained staff only.
- -20 °C
- Storage Comment
- Store at -20°C for one year. Avoid repeated freeze/thaw cycles.
- Expiry Date
- 12 months
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- Caspase 8 (CASP8)
- Alternative Name
- Caspase 8 (CASP8 Products)
- casp8-A antibody, xCaspase-8 antibody, casp8 antibody, CASP8 antibody, CASP-8 antibody, FLICE antibody, MACH antibody, Mch5 antibody, ALPS2B antibody, CAP4 antibody, Casp-8 antibody, MCH5 antibody, caspase-8 antibody, zgc:92075 antibody, caspase 8 L homeolog antibody, death related ced-3/Nedd2-like protein antibody, caspase 8 antibody, caspase-8 antibody, caspase-8-like antibody, caspase 8, apoptosis-related cysteine peptidase antibody, casp8.L antibody, LOC661095 antibody, CASP8 antibody, LOC5563818 antibody, casp8 antibody, LOC100222284 antibody, Casp8 antibody
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 CASP1. 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.
Subcellular location: Cytoplasm
Synonyms: MACH, Mch5, FLICE, CASP-8, Caspase-8, Casp8
- Gene ID
- Apoptosis, Caspase Cascade in Apoptosis, TLR Signaling, Activation of Innate immune Response, Tube Formation, Positive Regulation of Endopeptidase Activity, Toll-Like Receptors Cascades