Overview

  • Product nameAnti-Caspase-8 antibody
    See all Caspase-8 primary antibodies
  • Description
    Rabbit polyclonal to Caspase-8
  • Specificityab25901 recognises the p18 form of Caspase-8.
  • Tested applicationsSuitable for: ICC/IF, IHC-P, ICC, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide corresponding to Human Caspase-8 (C terminal). Synthetic peptide (Human) - a 16 amino acid peptide from near the carboxy-terminus of human Caspase-8 isoform A.
    Database link: Q14790

  • Positive control
    • Jurkat cell lysate IHC-P: Human spleen FFPE tissue sections.

Properties

Applications

Our Abpromise guarantee covers the use of ab25901 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF Use a concentration of 20 µg/ml.
IHC-P Use a concentration of 1 - 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC Use a concentration of 2 µg/ml.
WB Use a concentration of 0.5 - 2 µg/ml. Detects a band of approximately 18 kDa (predicted molecular weight: 18 kDa).

Target

  • FunctionMost upstream protease of the activation cascade of caspases responsible for the TNFRSF6/FAS mediated and TNFRSF1A induced cell death. Binding to the adapter molecule FADD recruits it to either receptor. The resulting aggregate called death-inducing signaling complex (DISC) performs CASP8 proteolytic activation. The active dimeric enzyme is then liberated from the DISC and free to activate downstream apoptotic proteases. Proteolytic fragments of the N-terminal propeptide (termed CAP3, CAP5 and CAP6) are likely retained in the DISC. Cleaves and activates CASP3, CASP4, CASP6, CASP7, CASP9 and CASP10. May participate in the GZMB apoptotic pathways. Cleaves ADPRT. Hydrolyzes the small-molecule substrate, Ac-Asp-Glu-Val-Asp-
    -AMC. Likely target for the cowpox virus CRMA death inhibitory protein. Isoform 5, isoform 6, isoform 7 and isoform 8 lack the catalytic site and may interfere with the pro-apoptotic activity of the complex.
  • Tissue specificityIsoform 1, isoform 5 and isoform 7 are expressed in a wide variety of tissues. Highest expression in peripheral blood leukocytes, spleen, thymus and liver. Barely detectable in brain, testis and skeletal muscle.
  • Involvement in diseaseDefects in CASP8 are the cause of caspase-8 deficiency (CASP8D) [MIM:607271]. CASP8D is a disorder resembling autoimmune lymphoproliferative syndrome (ALPS). It is characterized by lymphadenopathy, splenomegaly, and defective CD95-induced apoptosis of peripheral blood lymphocytes (PBLs). It leads to defects in activation of T-lymphocytes, B-lymphocytes, and natural killer cells leading to immunodeficiency characterized by recurrent sinopulmonary and herpes simplex virus infections and poor responses to immunization.
  • Sequence similaritiesBelongs to the peptidase C14A family.
    Contains 2 DED (death effector) domains.
  • DomainIsoform 9 contains a N-terminal extension that is required for interaction with the BCAP31 complex.
  • Post-translational
    modifications
    Generation of the subunits requires association with the death-inducing signaling complex (DISC), whereas additional processing is likely due to the autocatalytic activity of the activated protease. GZMB and CASP10 can be involved in these processing events.
    Phosphorylated upon DNA damage, probably by ATM or ATR.
  • Cellular localizationCytoplasm.
  • Information by UniProt
  • Database links
  • Alternative names
    • ALPS2B antibody
    • Amyotrophic lateral sclerosis 2 chromosomal region candidate gene 12 protein antibody
    • Apoptotic cysteine protease antibody
    • Apoptotic protease Mch-5 antibody
    • Apoptotic protease Mch5 antibody
    • CAP4 antibody
    • CASP-8 antibody
    • CASP8 antibody
    • CASP8_HUMAN antibody
    • Caspase 8 antibody
    • Caspase 8 apoptosis related cysteine peptidase antibody
    • Caspase-8 subunit p10 antibody
    • CED 3 antibody
    • FADD Like ICE antibody
    • FADD-homologous ICE/CED-3-like protease antibody
    • FADD-like ICE antibody
    • FLICE antibody
    • FLJ17672 antibody
    • ICE-like apoptotic protease 5 antibody
    • MACH alpha 1/2/3 protein antibody
    • MACH antibody
    • MACH beta 1/2/3/4 protein antibody
    • MCH5 antibody
    • MGC78473 antibody
    • MORT1 associated ced 3 homolog antibody
    • MORT1-associated CED-3 homolog antibody
    • OTTHUMP00000163717 antibody
    • OTTHUMP00000163720 antibody
    • OTTHUMP00000163724 antibody
    • OTTHUMP00000163725 antibody
    • OTTHUMP00000165062 antibody
    • OTTHUMP00000165063 antibody
    • OTTHUMP00000165064 antibody
    • OTTHUMP00000206552 antibody
    • OTTHUMP00000206582 antibody
    see all

Anti-Caspase-8 antibody images

  • Immunohistochemistry of Caspase-8 in human spleen tissue with Caspase-8 antibody at 5 µg/ml.

  • All lanes : Anti-Caspase-8 antibody (ab25901) at 1 µg/ml

    Lane 1 : Jurkat cell lysates with Absence of blocking buffer
    Lane 2 : Jurkat cell lysates with Presence of blocking buffer


    Predicted band size : 18 kDa
  • Lane 1 : Anti-Caspase-8 antibody (ab25901) at 0.5 µg/ml
    Lane 2 : Anti-Caspase-8 antibody (ab25901) at 1 µg/ml
    Lane 3 : Anti-Caspase-8 antibody (ab25901) at 2 µg/ml

    Lane 1 : Jurkat cell lysate.
    Lane 2 : Jurkat cell lysate.
    Lane 3 : Jurkat cell lysate.


    Predicted band size : 18 kDa
    Observed band size : 20 kDa (why is the actual band size different from the predicted?)
  • Anti-Caspase-8 antibody (ab25901) at 1 µg/ml + Recombinant human Caspase-8 protein (ab54117) at 0.01 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 18 kDa


    Exposure time : 1 minute
  • ab25901 at 2ug/ml staining Caspase-8 in Jurkat cells by ICC/IF

  • IHC image of Caspase-8 staining in human spleen formalin fixed paraffin embedded tissue section, performed on a Leica Bond system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab25901, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

References for Anti-Caspase-8 antibody (ab25901)

This product has been referenced in:
  • Kauerova T  et al. Antiproliferative and Pro-Apoptotic Effect of Novel Nitro-Substituted Hydroxynaphthanilides on Human Cancer Cell Lines. Int J Mol Sci 17:N/A (2016). Read more (PubMed: 27483236) »
  • Zhong R  et al. Kuntai Capsule Inhibited Endometriosis via Inducing Apoptosis in a Rat Model. Evid Based Complement Alternat Med 2016:5649169 (2016). Rat . Read more (PubMed: 27597876) »

See all 11 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (stomach cancer, colon cancer)
Loading amount 30 µg
Specification stomach cancer, colon cancer
Treatment 10uM canabinoid antagonist for 24hrs
Gel Running Conditions Reduced Denaturing (8% SDS-PAGE)
Blocking step Milk as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Apr 05 2013

Thanks for your call today and for letting us know about the trouble analyzing the results of ab25901 in Western blot.

As w discussed, I'm sending a vial of ab119892 free of charge on the order ***, which should arrive on Monday. Please keep ...

Read More

Thank you for contacting us. The image on our datasheet was generated using 15ug of Jurkat cell lysate per lane under reduced and denaturing conditions. The membrane was blocked for 2 hours with 5% milk. The antibody was ab25901 was diluted 0.5- ...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Rat Cell lysate - whole cell (Rat Type II Cell)
Loading amount 20 µg
Specification Rat Type II Cell
Gel Running Conditions Reduced Denaturing (8-16% Tris-Glycine)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Aug 28 2009

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"