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Human/Mouse Caspase-8 Antibody

Catalog #: AF1650
21 Citations Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
AF1650
AF1650-SP
Detection of Human Caspase‑8 by Western Blot.
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Product Details
Citations (21)
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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 Detection of Human Caspase-8 antibody by Western Blot. View Larger

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 Detection of Human Caspase-8 antibody by Simple Western<SUP>TM</SUP>. View Larger

Detection of Human Caspase‑8 by Simple WesternTM. 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.

Western Blot Detection of Human Caspase-8 by Western Blot View Larger

Detection of Human Caspase-8 by Western Blot Western blot analysis of the expression of a number of proteins impacting proliferation and survival of ovarian cancer cells 48 hours after alteration of the Spry1 expression. Results indicate activation of apoptotic processes detected as overexpression of the pro-apoptotic Bax, decreased expression of the antiapoptotic proteins Bcl-2 and Bcl-xl, attenuation of procaspases 3, 7, 8 and 9 and cleavage of PARP in the Spry1-transfected SKOV-3 cells (left). Moreover, reduced expression of the activated forms of ERK and AKT along with increased expression of PTEN with concomitant decrease of phospho-PTEN (right) implicates repression of ERK as well as of AKT, with involvement of PTEN for the latter, in Spry1-induced inhibition of cell proliferation and survival. No significant change in the expression pattern of these proteins was found in the Spry1-silenced 1A9 cells. Image collected and cropped by CiteAb from the following publication (https://ovarianresearch.biomedcentral.com/articles/10.1186/1757-2215-7-61), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
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Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
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-FLIPL 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-FLIPL 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-FLIPL, 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-FLIPL, c-FLIPL 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
  1. Chowdhury, I. et al. (2008) Comp. Biochem. Physiol. B 151:10.
  2. Boatright, K.M. & G.S. Salvesen (2003) Curr. Opin. Cell Biol. 15:725.
  3. Launay, S. et al. (2005) Oncogene 24:5137.
  4. Srinivasula, S.M. et al. (1996) Proc. Natl. Acad. Sci. USA 93:14486.
  5. Hughes, M.A. et al. (2009) Mol. Cell 35:265.
  6. Lamkanfi, M. et al. (2007) Cell Death Differ. 14:44.
  7. Fernandes-Alnemri, T. et al. (1996) Proc. Natl. Acad. Sci. USA 93:7464.
  8. Boldin, M.P. et al. (1996) Cell 85:803.
  9. Muzio, M. et al. (1996) Cell 85:817.
  10. Donepudi, M. et al. (2003) Mol. Cell 11:543.
  11. Boatright, K.M. et al. (2003) Mol. Cell 11:529.
  12. Golks, A. et al. (2006) Cell Death Differ. 13:489.
  13. Scaffidi, C. et al. (1997) J. Biol. Chem. 272:26953.
  14. 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

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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.

21 Citations: Showing 1 - 10
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  1. Pentamidine Blocks Hepatotoxic Injury in Mice
    Authors: E Zhao, G Ilyas, F Cingolani, JH Choi, F Ravenelle, KE Tanaka, MJ Czaja
    Hepatology, 2017-07-20;0(0):.
  2. Autophagy promotes T-cell survival through degradation of proteins of the cell death machinery.
    Authors: Kovacs JR, Li C, Yang Q et al.
    Cell Death Differ.
  3. 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, 2022-11-24;0(0):.
    Species: Complex Species Category
    Sample Types: Cell Lysates
    Applications: Western Blot
  4. 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, 2021-08-20;0(0):.
    Species: Mouse
    Sample Types: Cell Lysates
    Applications: Western Blot
  5. 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, 2021-01-12;0(0):.
    Species: Mouse
    Sample Types: Tissue Homogenates
    Applications: Western Blot
  6. 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, 2020-10-02;11(10):822.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  7. Gasdermin pores permeabilize mitochondria to augment caspase-3 activation during apoptosis and inflammasome activation
    Authors: Corey Rogers, Dan A. Erkes, Alexandria Nardone, Andrew E. Aplin, Teresa Fernandes-Alnemri, Emad S. Alnemri
    Nature Communications
  8. Bryostatin and its synthetic analog, picolog rescue dermal fibroblasts from prolonged stress and contribute to survival and rejuvenation of human skin equivalents
    Authors: Tapan K. Khan, Paul A. Wender, Daniel L. Alkon
    Journal of Cellular Physiology
  9. 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., 2017-06-01;39(6):1010428317711.
    Species: Xenograft
    Sample Types: Cell Lysates
    Applications: Western Blot
  10. Ligand stimulation of CD95 induces activation of Plk3 followed by phosphorylation of caspase-8
    Authors: Christina Helmke, Monika Raab, Franz Rödel, Yves Matthess, Thomas Oellerich, Ranadip Mandal et al.
    Cell Research
  11. 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, 2014-06-08;7(0):61.
    Species: Human
    Sample Types: Whole Cells
    Applications: Western Blot
  12. 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, 2014-05-22;5(0):3920.
    Species: Mouse
    Sample Types: Whole Tissue
    Applications: IHC
  13. TolC-Dependent Modulation of Host Cell Death by the Francisella tularensis Live Vaccine Strain
    Authors: Christopher R. Doyle, Ji-An Pan, Patricio Mena, Wei-Xing Zong, David G. Thanassi
    Infection and Immunity
  14. Propranolol, a beta -adrenoceptor antagonist, worsens liver injury in a model of non-alcoholic steatohepatitis
    Authors: Chad McKee, Junpei Soeda, Esra Asilmaz, Barbara Sigalla, Maelle Morgan, Nicoletta Sinelli et al.
    Biochemical and Biophysical Research Communications
  15. 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, 2013-01-26;25(5):1245-51.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  16. 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, 2012-12-13;288(5):2952-63.
    Species: Mouse
    Sample Types: Cell Lysates
    Applications: Western Blot
  17. 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., 2011-05-31;31(15):3158-70.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  18. TNF alpha Cooperates with IFN-gamma to Repress Bcl-xL Expression to Sensitize Metastatic Colon Carcinoma Cells to TRAIL-mediated Apoptosis
    Authors: Feiyan Liu, Xiaolin Hu, Mary Zimmerman, Jennifer L. Waller, Ping Wu, Andrea Hayes-Jordan et al.
    PLoS ONE
  19. 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., 2008-02-20;82(9):4502-10.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Neutralization
  20. Amiodarone as an autophagy promoter reduces liver injury and enhances liver regeneration and survival in mice after partial hepatectomy.
    Authors: Lin CW, Chen YS, Lin CC et al.
    Sci Rep.
  21. A Novel Anticancer Agent, 8-Methoxypyrimido[4',5':4,5]thieno(2,3-) Quinoline-4(3H)-One Induces Neuro 2a Neuroblastoma Cell Death through p53-Dependent, Caspase-Dependent and -Independent Apoptotic Pathways.
    Authors: Sahu Upasana, Sidhar Himakshi, Ghate Pankaj S et al.
    PLoS One.

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