Home » Caspase-8 » Caspase 8 Research Products » Caspase-8 Antibody [Unconjugated]

Caspase-8 Antibody [Unconjugated]

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Western blot shows lysates of Jurkat human leukemic T cell line and DA3 mouse myeloma cell line treated with 1 µM staurosporine for the indicated time. PVDF membrane was probed with 0.5 µg/mL Goat Anti-Human/Mouse Caspase‑8 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF705) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). Specific bands for Caspase‑8 precursor were detected at approximately 60 kDa (as indicated in upper panal) and specific bands for cleaved Caspase-8 were detected at approximately 14-18 kDa (as indicated in lower panal). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 4.
Western blot shows lysates of Jurkat human leukemic T cell line and DA3 mouse myeloma cell line treated with 1 µM staurosporine for the indicated time. PVDF membrane was probed with 0.5 µg/mL Goat Anti-Human/Mouse ...read more
Genetic Strategies: Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and Caspase-8 knockout HeLa cell line (KO). PVDF membrane was probed with 0.5 µg/mL of Goat ...read more
Simple Western lane view shows lysates of Jurkat human acute T cell leukemia cell line untreated or treated with Staurosporine (STS) for 2 hours, loaded at 0.2 mg/mL. Specific bands were detected for Caspase‑8 ...read more

Product Details

Summary
Reactivity Hu, MuSpecies Glossary
Applications WB, Simple Western, KO
Clonality
Polyclonal
Host
Goat
Conjugate
Unconjugated
Concentration
LYOPH
Validated by:
     

Genetic Strategies

    

Order Details

View Available Formulations
Catalog# & Formulation Size Price
Carrier Free
AF705-SP
100% Guarantee on every product

Caspase-8 Antibody [Unconjugated] Summary

Immunogen
E. coli-derived recombinant human Caspase-8
Ser234-Asp496
Accession # AAC50645
Specificity
Detects human and mouse Caspase-8 in Western blots.
Source
N/A
Isotype
IgG
Clonality
Polyclonal
Host
Goat
Gene
CASP8
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.
Learn about the Innovator's Reward

Applications/Dilutions

Dilutions
  • Western Blot 0.5 ug/mL
  • Simple Western 5 ug/mL
  • Knockout Validated
Control
Caspase-8 Knockout HeLa Cell Lysate
Reviewed Applications
Read 1 Review rated 3
using
AF705 in the following applications:
Publications
Read Publications using
AF705 in the following applications:

Packaging, Storage & Formulations

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.
Buffer
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
Preservative
No Preservative
Concentration
LYOPH
Reconstitution Instructions
Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Caspase-8 Antibody [Unconjugated]

  • 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

Background

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).

  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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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Caspase-8 Antibody
[Unconjugated]
AF705
Caspase-8 Antibody [Unconjugated]
Species: Hu, Mu
Applications: WB, Simple Western, KO

Publications for Caspase-8 Antibody (AF705)(5)

We have publications tested in 2 confirmed species: Human, Mouse.

We have publications tested in 2 applications: ICC, Western Blot.


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(1)
Western Blot
(4)
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Review for Caspase-8 Antibody (AF705) (1) 31

Average Rating: 3
(Based on 1 review)
We have 1 review tested in 1 species: Human.

Reviews using AF705:
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WB
(1)
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Human
(1)
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Images Ratings Applications Species Date Details
 
reviewed by:
Anonymous
 WB Human 01/09/2015
View

Summary

ApplicationWestern Blot
Sample TestedSee PMID 20074295
SpeciesHuman

Blocking

Blocking DetailsSee PMID 20074295

Details

Detection NotesSee PMID 20074295

Comments

CommentsPublished: See PMID 20074295Independent review from antibodyresource.com

Product General Protocols

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

Video Protocols

WB Video Protocol

FAQs for Caspase-8 Antibody (AF705). (Showing 1 - 1 of 1 FAQs).

  1. May we ask the MW difference between the active form and the proform of Caspase 8 while performing WB?
    • In treated cells induced to undergo apoptosis, caspase-8 migrates as a 55/53 kDa (pro-form), 41/42 kDa (a cleaved/active or intermediate form), and 18 kDa (active form). The proform (55/53 kDa) is still seen in treated cells because not all cells undergo apoptosis at once.

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Secondary Antibodies

 

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Blogs on Caspase-8.

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Caspase 3, the executioner of apoptosis
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Caspase-8 - a pro-apoptotic protein with dynamic roles in normal physiology and pathology
Caspases are a family of cysteine-aspartic acid proteases that are responsible for the initiation and execution of apoptosis. Caspase-8 is a 55 kDa protein expressed as an inactive procaspase that resides in the cytosol. Activation of Caspase-8 req...  Read full blog post.
Caspase 8 - a key mediator of apoptosis
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210-TA-005
Caspase-8 Activity Assay Kit (Colorimetric) [NBP2-54817] - The assay was performed on murine insulinoma cell line following Novu's protocol.
Caspase-8 Activity Assay Kit (Colorimetric)
NBP2-54817-100Assays

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 Anonymous
01/09/2015
Application: WB
Species: Human
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Bioinformatics

Gene Symbol CASP8
Uniprot