Human/Mouse Caspase-8 Antibody

Catalog #: AF1650
11 Citations Datasheet / COA / SDS

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AF1650
AF1650-SP

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Detection of Human Caspase‑8 by Western Blot.

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Citations (11)

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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human/Mouse Caspase-8 Antibody Summary

Species Reactivity

Human, Mouse

Specificity

Detects human/mouse Caspase-8 and cleavage products. Detects multiple isoforms of Caspase-8.

Source

Polyclonal Rabbit IgG

Purification

Antigen Affinity-purified

Immunogen

E. coli-derived recombinant human Caspase-8
Ser217-Asp384 (Asp285His) (p18 subunit), Leu385-Asp479 (p10 subunit)
Accession # Q14790

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Western Blot

0.5 µg/mL

See below

Simple Western

5 µg/mL

See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot

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Detection of Human Caspase‑8 by Western Blot. Western blot shows lysates of Jurkat human acute T cell leukemia cell line untreated (-) or treated (+) with 1 mM staurosporine (STS) for for 3 hours. PVDF membrane was probed with 0.5 µg/mL of Rabbit Anti-Human/Mouse Caspase-8 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1650), followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). For additional reference Recombinant Human Caspase-8 (Catalog # 705-C8) was included. Specific bands were detected for Caspase-8 precursor at approximately 57-60 kDa (as indicated). In STS-treated samples, specific bands were detected for Caspase-8 p41/43 subunit at approximately 41 and 43 kDa (as indicated) and Caspase-8 p18, p14, and p10 subunits at approximately 18 kDa, 14 kDa, and 10 kDa, respectively (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 4.

Simple Western

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Detection of Human Caspase‑8 by Simple Western^TM. Simple Western lane view shows lysates of Jurkat human acute T cell leukemia cell line untreated (-) or treated (+) with 1 mM Staurosporine (STS) for 3 hours, loaded at 0.2 mg/mL. Specific bands were detected for Caspase‑8 at approximately 58-62 kDa (as indicated) using 5 µg/mL of Rabbit Anti-Human/Mouse Caspase‑8 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1650). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS.

Reconstitution Buffer Available

Reconstitution Buffer 1 (PBS)

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: Caspase-8

Caspase-8 (Cysteine-aspartic acid protease 8/Casp8a; also named MCH5, FLICA and MACH alpha 1) is a 28 kDa member of the peptidase C14A family of enzymes (1, 2, 3). It is widely expressed and is considered an initiating caspase for the apoptotic cascade (4). Caspase-8 acts on a wide variety of substrates, including procaspases-3, 4, 6, 7, 9 and 10, c-FLIP_L and procaspase-8 itself (1, 5, 6). Human procaspase-8a is a 54‑56 kDa, 479 amino acid (aa) protein (4, 7, 8, 9). It contains two N-terminal death domains (aa 1‑177), followed by a catalytic site that utilizes His317Gly318 plus Cys360. Normally, it is an inactive, cytosolic monomer (1, 10, 11). But following death-domain (DD) containing receptor oligomerization, Caspase-8 is recruited to the death-inducing signaling complex (DISC) that forms around the death domains of the oligomerized receptor (12). FADD/CAP-1 is recruited first, followed by procaspase-8/CAP-4 and, possibly, c-FLIP_L and procaspase-10 (12). The recruitment, or concentration, of procaspase-8 induces homodimerization. This act alone is sufficient for activation. However, the activity level is modest at best, and appears to be directed towards either itself, or c-FLIP_L, which is known to form a functional heterodimer with procaspase-8 (5, 11). When directed towards itself, autocleavage occurs first between Asp374Ser375, generating a 43 kDa (p43) N-terminal (aa 1‑374) and an 11 kDa C-terminal (aa 375 - 479) fragment. The C-terminus is further cleaved between Asp384Leu385 to generate a mature p10 subunit (aa 385‑479). The p43 subunit is next cleaved twice, once between Asp216Ser217, and again between Asp210Ser211 to generate a 26 kDa DD-containing prodomain (aa 1‑210) with an additional 18 kDa mature p18 subunit (aa 217‑374) (12). p18 and p10 noncovalently associate to form a 28 kDa heterodimer, which subsequently associates with another p18:p10 heterodimer to form an active, mature Caspase-8 molecule. This leaves the DISC to act on downstream apoptotic procaspases. In the event procaspase-8 comes to the DISC complexed with c-FLIP_L, c-FLIP_L will be cleaved by procaspase-8, generating a p43 fragment that is analogous to the Caspase-8 p43 subunit. This fragment, however, appears not to be an intermediate in a proteolytic cascade. Rather, it serves as a functional subunit, interacting with TRAF2 and activating NF kappa B. This may account for many of the nonapoptotic activities associated with Caspase-8 (5, 6, 13). Mature human and mouse Caspase-8a heterodimers are 73% aa identical (14).

References

Chowdhury, I. et al. (2008) Comp. Biochem. Physiol. B 151:10.
Boatright, K.M. & G.S. Salvesen (2003) Curr. Opin. Cell Biol. 15:725.
Launay, S. et al. (2005) Oncogene 24:5137.
Srinivasula, S.M. et al. (1996) Proc. Natl. Acad. Sci. USA 93:14486.
Hughes, M.A. et al. (2009) Mol. Cell 35:265.
Lamkanfi, M. et al. (2007) Cell Death Differ. 14:44.
Fernandes-Alnemri, T. et al. (1996) Proc. Natl. Acad. Sci. USA 93:7464.
Boldin, M.P. et al. (1996) Cell 85:803.
Muzio, M. et al. (1996) Cell 85:817.
Donepudi, M. et al. (2003) Mol. Cell 11:543.
Boatright, K.M. et al. (2003) Mol. Cell 11:529.
Golks, A. et al. (2006) Cell Death Differ. 13:489.
Scaffidi, C. et al. (1997) J. Biol. Chem. 272:26953.
Sakamaki, K. et al. (1998) Eur. J. Biochem. 253:399.

Entrez Gene IDs

841 (Human); 12370 (Mouse)

Alternate Names

AIS; androgen receptor; CASP8; Caspase8; Caspase-8; DHTRTFM; Dihydrotestosterone receptorHYSP1; HUMARA; Mch5; NR3C4KD; Nuclear receptor subfamily 3 group C member 4; SMAX1SBMA; spinal and bulbar muscular atrophy

Product Datasheets

Product Datasheet

COA

SDS

Related Research Areas

Citations for Human/Mouse Caspase-8 Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

11 Citations: Showing 1 - 10
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The expression of the Sprouty 1 protein inversely correlates with growth, proliferation, migration and invasion of ovarian cancer cells.
Authors: Masoumi-Moghaddam, Samar, Amini, Afshin, Ehteda, Anahid, Wei, Ai-Qun, Morris, David La
J Ovarian Res, 2104;7(0):61.
Species: Human
Sample Types: Whole Cells
Applications: Western Blot
Caspase-8 is required for HSV-1-induced apoptosis and promotes effective viral particle release via autophagy inhibition
Authors: F Marino-Mer, A Klett, E Papaianni, SFA Drago, B Macchi, MG Rincón, F Andreola, A Serafino, S Grelli, A Mastino, C Borner
Cell Death and Differentiation, 2022;0(0):.
Species: Complex Species Category
Sample Types: Cell Lysates
Applications: Western Blot
Targeting Cpt1a-Bcl-2 interaction modulates apoptosis resistance and fibrotic remodeling
Authors: L Gu, R Surolia, JL Larson-Cas, C He, D Davis, J Kang, VB Antony, AB Carter
Cell Death and Differentiation, 2021;0(0):.
Species: Mouse
Sample Types: Cell Lysates
Applications: Western Blot
Complement-5 inhibition deters progression of fulminant hepatitis to acute liver failure in murine models
Authors: J Kusakabe, K Hata, H Miyauchi, T Tajima, Y Wang, I Tamaki, J Kawasoe, Y Okamura, X Zhao, T Okamoto, T Tsuruyama, S Uemoto
Cellular and Molecular Gastroenterology and Hepatology, 2021;0(0):.
Species: Mouse
Sample Types: Tissue Homogenates
Applications: Western Blot
AFP promotes HCC progression by suppressing the HuR-mediated Fas/FADD apoptotic pathway
Authors: T Chen, X Dai, J Dai, C Ding, Z Zhang, Z Lin, J Hu, M Lu, Z Wang, Y Qi, L Zhang, R Pan, Z Zhao, L Lu, W Liao, X Lu
Cell Death Dis, 2020;11(10):822.
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot
Ferrichrome identified from Lactobacillus casei ATCC334 induces apoptosis through its iron-binding site in gastric cancer cells
Authors: M Ijiri, M Fujiya, H Konishi, H Tanaka, N Ueno, S Kashima, K Moriichi, J Sasajima, K Ikuta, T Okumura
Tumour Biol., 2017;39(6):1010428317711.
Species: Xenograft
Sample Types: Cell Lysates
Applications: Western Blot
NRBF2 regulates autophagy and prevents liver injury by modulating Atg14L-linked phosphatidylinositol-3 kinase III activity.
Authors: Lu, Jiahong, He, Liqiang, Behrends, Christia, Araki, Masatake, Araki, Kimi, Jun Wang, Qing, Catanzaro, Joseph M, Friedman, Scott L, Zong, Wei-Xing, Fiel, M Isabel, Li, Min, Yue, Zhenyu
Nat Commun, 2014;5(0):3920.
Species: Mouse
Sample Types: Whole Tissue
Applications: IHC
TCEA3 binds to TGF-beta receptor I and induces Smad-independent, JNK-dependent apoptosis in ovarian cancer cells.
Authors: Cha Y, Kim D, Hyun J, Kim S, Park K
Cell Signal, 2013;25(5):1245-51.
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot
Hyperactivation of the mammalian degenerin MDEG promotes caspase-8 activation and apoptosis.
Authors: Pan, Ji-An, Fan, Yongjun, Gandhirajan, Rajesh K, Madesh, Muniswam, Zong, Wei-Xing
J Biol Chem, 2013;288(5):2952-63.
Species: Mouse
Sample Types: Cell Lysates
Applications: Western Blot
Inhibition of protein degradation induces apoptosis through a microtubule-associated protein 1 light chain 3-mediated activation of caspase-8 at intracellular membranes.
Authors: Pan JA, Ullman E, Dou Z
Mol. Cell. Biol., 2011;31(15):3158-70.
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot

Theiler's virus-induced intrinsic apoptosis in M1-D macrophages is Bax mediated and restricts virus infectivity: a mechanism for persistence of a cytolytic virus.
Authors: Son KN, Becker RP, Kallio P, Lipton HL
J. Virol., 2008;82(9):4502-10.
Species: Mouse
Sample Types: Whole Cells
Applications: Neutralization