Product Pathways - Apoptosis
Cleaved Caspase-8 (Asp387) (D5B2) XP® Rabbit mAb (Mouse Specific) #8592
PhosphoSitePlus® protein, site, and accession data: Casp8
| Applications | Reactivity | Sensitivity | MW (kDa) | Isotype |
|---|---|---|---|---|
| W IP IF-IC F | M | Endogenous | 18, 43 | Rabbit IgG |
Applications Key:
W=Western Blotting
IP=Immunoprecipitation
IF-IC=Immunofluorescence (Immunocytochemistry)
F=Flow Cytometry
Reactivity Key:
M=Mouse
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.
Protocols
Specificity / Sensitivity
Cleaved Caspase-8 (Asp387) (D5B2) XP® Rabbit mAb (Mouse Specific) recognizes endogenous levels of mouse caspase-8 only when cleaved at Asp387. In addition to the p18 subunit, this antibody also detects cleavage products containing the pro-domain with the p18 subunit (p43).
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Asp387 of mouse caspase-8 protein.
Western Blotting
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct overexpressing mouse caspase-8 (mCasp8) (+), using Cleaved Caspase-8 (Asp387) (D5B2) XP® Rabbit mAb (Mouse Specific).
Western Blotting
Western blot analysis of extracts from CTLL-2 cells, untreated or treated with cycloheximide (CHX, 10 μg/ml, overnight) followed by hTNF-α #8902 (20 ng/ml, 4 hours), using Cleaved Caspase-8 (Asp387) (D5B2) XP® Rabbit mAb (Mouse Specific).
Flow Cytometry
Flow cytometric analysis of CTLL-2 cells, untreated (red) or treated with cyclohexamide and hTNF-α #8902 (blue), using Cleaved Caspase-8 (Asp387) (D5B2) XP® Rabbit mAb (Mouse Specific).
IF-IC
Confocal immunofluorescent analysis of Raw 264.7 cells, untreated (left) or treated with TNF-α and Cycloheximide (20 ng/ml and 2.5 ug/ml, 4 hours and 3 hours; right), using Cleaved Caspase-8 (Asp387) (D5B2) XP® Rabbit mAb (Mouse Specific) (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Background
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.
- Muzio, M. et al. (1996) Cell 85, 817-827.
- Boldin, M.P. et al. (1996) Cell 85, 803-815.
- Fernandes-Alnemri, T. et al. (1996) Proc. Natl. Acad. Sci. USA 93, 7464-7469.
Application References
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For Research Use Only. Not For Use In Diagnostic Procedures.
