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Purified Mouse Anti-Human Caspase-8
Purified Mouse Anti-Human Caspase-8
Western blot analysis of caspase-8. Lysates from control (lanes 1-3) and camptothecin treated Jurkat cells (lanes 4-6) were probed with anti-human caspase-8 (clone 3-1-9, Cat. No. 551243) at the following concentrations: 0.5 (lanes 1,4), 0.25 (lanes 2,5), and 0.125 µg/ml (lanes 3,6). Caspase-8 is identified as 55/50 kDa (proform), 40/36 kDa (cleaved), and 23 kDa (cleaved) bands in treated cells and the 55 kDa band in control cells.
Purified Mouse Anti-Human Caspase-8
(+)=positive, (-)=negative, (NT)=not tested
Purified Mouse Anti-Human Caspase-8
Immunoprecipitation/western blot analysis of caspase-8. Lysates from control (lane 1) or camptothecin-treated Jurkat cells (lane 2) were each immunoprecipitated with anti-human caspase-8 (clone 3-1-9), and western blotted with anti-human caspase-8 (clone 3-1-9). The 55 kDa proform caspase-8 was identified in both control and camptothecin-treated cells.
Western blot analysis of caspase-8. Lysates from control (lanes 1-3) and camptothecin treated Jurkat cells (lanes 4-6) were probed with anti-human caspase-8 (clone 3-1-9, Cat. No. 551243) at the following concentrations: 0.5 (lanes 1,4), 0.25 (lanes 2,5), and 0.125 µg/ml (lanes 3,6). Caspase-8 is identified as 55/50 kDa (proform), 40/36 kDa (cleaved), and 23 kDa (cleaved) bands in treated cells and the 55 kDa band in control cells.
(+)=positive, (-)=negative, (NT)=not tested
Immunoprecipitation/western blot analysis of caspase-8. Lysates from control (lane 1) or camptothecin-treated Jurkat cells (lane 2) were each immunoprecipitated with anti-human caspase-8 (clone 3-1-9), and western blotted with anti-human caspase-8 (clone 3-1-9). The 55 kDa proform caspase-8 was identified in both control and camptothecin-treated cells.
Product Details
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BD Pharmingen™
FLICE, MACH-1, Mch5
Human (QC Testing)
Mouse IgG1, κ
Human Caspase-8 full-length recombinant protein
Western blot (Routinely Tested), Immunoprecipitation (Tested During Development)
55/50 kDa, 40/36 kDa, 23 kDa
0.5 mg/ml
AB_394115
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. Store undiluted at 4°C.

Recommended Assay Procedures

Applications include western blot analysis ( 0.125 - 0.5 µg /ml) and immunoprecipitation (4 µg /200µg cell lysate ). Jurkat T cells ( ATCC CRL-1573) are suggested as a positive control. BD Biosciences Pharmingen offers several caspase-8 antibodies. A Jurkat model cell system was used to evaluate these antibodies; these results are summarized in the following table. However, actual bands observed could vary according to the cell model system or treatment used.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  3. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
551242 Rev. 1
Antibody Details
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3-1-9

Caspase-8 (FLICE/MACH-1) is a 55 kDa cytosolic protein with homology to the CD95/Fas-associated signal transducer, FADD/MORT-1, as well as to other caspase (ICE/Ced-3) cysteine proteases. The N-terminal region of caspase-8 contains an amino acid sequence, termed the death domain, that facilitates caspase-8-FADD direct interaction. FADD therefore acts as an adapter molecule, allowing caspase-8 to become recruited to the cytoplasmic region of Fas following receptor activation. Viral proteins (v-FLIPS) which inhibit recruitment and activation of caspase-8 have been isolated. Caspase-8 is produced as a proenzyme (55/50 kDa doublet) which upon receptor aggregation is proteolytically cleaved into smaller subunits of 40/36 (doublet), and 23 kDa. Overexpression of caspase-8 is sufficient to induce apoptosis in certain cell lines (e.g., MCF-7) and this phenotype is blocked by overexpression of the caspase-3 protease inhibitor, CrmA. The antibody recognizes both the proform of human caspase-8 (55/50 kDa doublet) as well as the cleaved forms which migrate at 40/36 (doublet) and 23 kDa in SDS/PAGE. It also immunoprecipitates the full-length human caspase-8. Full-length recombinant human caspase-8 protein was used as immunogen.

551242 Rev. 1
Format Details
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Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
551242 Rev.1
Citations & References
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View product citations for antibody "551242" on CiteAb

Development References (4)

  1. Boesen-de Cock JG, Tepper AD, de Vries E, van Blitterswijk WJ, Borst J. Common regulation of apoptosis signaling induced by CD95 and the DNA-damaging stimuli etoposide and gamma-radiation downstream from caspase-8 activation. J Biol Chem. 1999; 274(20):14255-14261. (Biology). View Reference
  2. Cock JG, Tepper AD, de Vries E, van Blitterswijk WJ, Borst J. CD95 (Fas/APO-1) induces ceramide formation and apoptosis in the absence of a functional acid sphingomyelinase. J Biol Chem. 1998; 273(13):7560-7565. (Biology). View Reference
  3. Muzio M, Chinnaiyan AM, Kischkel FC, et al. FLICE, a novel FADD-homologous ICE/CED-3-like protease, is recruited to the CD95 (Fas/APO-1) death--inducing signaling complex. Cell. 1996; 85(6):817-827. (Biology).
  4. Thome M, Schneider P, Hofmann K, et al. Viral FLICE-inhibitory proteins (FLIPs) prevent apoptosis induced by death receptors. Nature. 1997; 386(6624):517-521. (Biology).
View All (4) View Less
551242 Rev. 1

 

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.