AdipoGen Life Sciences

anti-Caspase-8 (mouse), mAb (1G12)

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AG-20T-0137-C100100 µgCHF 350.00

Figure 1: Flow cytometry data in permeabilized cells of overexpressed FLAG-epitope-tagged pro-caspase-8 (active-site-mutant) in 293T cells using anti-caspase-8 mAbs (1G12) (Prod. No. AG-20T-0137), anti-FLAG or control. As a positive control staining with a MAb to FLAG was performed. As a negative control an isotype control was employed.

Figure 2: Western blot using anti-Caspase-8 (mouse), mAb (1G12) (Prod. No. AG-20T-0137) detecting endogenous caspase-8 in various mouse cell line, but not in human cell lines.

Figure 3: Western blot using anti-Caspase-8 (mouse), mAb (1G12) (Prod. No. AG-20T-0137) detecting endogenous caspase-8 in MEFs from WT mice, but not in MEFs from caspase-8-/- mice. Several smaller bands detected in the caspase-8-/- MEFs, correspond to truncated forms of caspase-8 made in the caspase-8-/- mice since only exons 1 and 2 of mouse caspase-8 were deleted in these knock-out mice and not the region encoding the p20 subunit. Note: Extra bands marked by * are only seen in lysates from caspase-8-/- MEFs and not in lysates from any WT cell lines or mouse WT tissue.

Figure 4: Western blot using anti-Caspase-8 (mouse), mAb (1G12) (Prod. No. AG-20T-0137) detecting the cleaved active p20 subunit of mouse caspase-8 in addition to the caspase-8 precursor, upon an apoptotic stimulus e.g. by cross-linked rhsFasL.

Specifications / Handling

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Product Details
Synonyms	Apoptotic Cysteine Protease; Apoptotic Protease Mch-5; CAP4; FADD-homologous ICE/ced-3-like Protease
Product Type	Monoclonal Antibody
Properties
Clone	1G12
Isotype	Rat IgG1κ
Immunogen/Antigen	Recombinant mouse caspase-8 (p20 subunit).
Application	ELISA: (recombinant mouse caspase-8) Flow Cytometry: (overexpressed in cell lines) Immunocytochemistry: (see literature reference 2) Western Blot: (excellent) Optimal conditions must be determined individually for each application.
Crossreactivity	Mouse
Specificity	Recognizes the p20 subunit of mouse caspase-8. Detects bands of ~55kDa (full-length caspase-8) and ~18kDa (apoptosis-induced cleavage fragment) by Western blot. Does not cross-react with human caspase-8.
Purity	≥95% (SDS-PAGE)
Purity Detail	Protein G-affinity purified (sepharose column).
Endotoxin Content	<0.1EU/μg purified protein (LAL test; Lonza).
Concentration	1mg/ml
Formulation	Liquid. In PBS containing 0.02% sodium azide.
Isotype Negative Control	Rat IgG1 Isotype Control
Shipping and Handling
Shipping	BLUE ICE
Short Term Storage	+4°C
Long Term Storage	-20°C
Handling Advice	After opening, prepare aliquots and store at -20°C. Avoid freeze/thaw cycles.
Use/Stability	Stable for at least 1 year after receipt when stored at -20°C.
Documents
MSDS	Download PDF
Product Specification Sheet
Datasheet	Download PDF

Product-specific References

Description

Procaspase-8 belongs to the family of caspases. Binding of FasL to Fas leads to formation of a receptor complex at the cellular membrane, which was named DISC. The DISC consists of oligomerized receptors, the DD-containing adaptor molecule FADD, procaspase-8, procaspase-10 and c-FLIP. The DISC structure provides a platform for the oligomerization of procaspase-8 that allows two procaspase-8 homodimers to be in the close proximity leading to the initial activation of procaspase-8. At the first cleavage step, the N-terminal p43/p41 and the C-terminal p30 cleavage products are generated. Importantly, these cleavage products already possess catalytic activity. At the second cleavage step, p43/p41 and p30 are processed to p10 and p20, respectively, which leads to the generation of the active caspase-8 heterotetramer (p20/p10)2.

Product References

Apaf-1 and caspase-9 do not act as tumor suppressors in myc-induced lymphomagenesis or mouse embryo fibroblast transformation: C.L. Scott, et al.; J. Cell Biol. 164, 89 (2004)
Modifications and intracellular trafficking of FADD/MORT1 and caspase-8 after stimulation of T lymphocytes: L.A. O'Reilly, et al.; Cell Death Differ. 11, 724 (2004)
Caspase-8 serves both apoptotic and nonapoptotic roles: T.B.Kang, et al.; J. Immunol. 73, 2976 (2004)
Role of membrane sphingomyelin and ceramide in platform formation for Fas-mediated apoptosis: M. Miyaji, et al.; J. Exp. Med. 202, 249 (2005)
Fatal Hepatitis Mediated by Tumor Necrosis Factor TNFα Requires Caspase-8 and Involves the BH3-Only Proteins Bid and Bim: T. Kaufmann, et al.; Immunity 30, 56 (2009)
Catalytic activity of the caspase-8-FLIPL complex inhibits RIPK3-dependent necrosis: A. Oberst, et al.; Nature 471, 363 (2011)
FLIP (Flice-like inhibitory protein) suppresses cytoplasmic double-stranded-RNA-induced apoptosis and NF-B and IRF3-mediated signaling: P. Handa, et al.; Cell Commun. Signal. 9, 16 (2011)
XIAP restricts TNF- and RIP3-dependent cell death and inflammasome activation: M. Yabal, et al.; Cell Rep. 7, 1796 (2014)

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