Overview

  • Product name
  • Description
    Rabbit polyclonal to Caspase-8
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IHC-P, IHC-Frmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Cow, Human, Monkey
  • Immunogen

    Recombinant full length protein corresponding to Human Caspase-8.
    Database link: Q14790

  • Positive control
    • Stomach.

Properties

Applications

Our Abpromise guarantee covers the use of ab4052 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
IHC-P 1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IHC-Fr 1/50 - 1/100.

Target

  • Function
    Most 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 specificity
    Isoform 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 disease
    Defects 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 similarities
    Belongs to the peptidase C14A family.
    Contains 2 DED (death effector) domains.
  • Domain
    Isoform 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 localization
    Cytoplasm.
  • 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

Images

  • ab4052 staining Caspase-8 in human kidney tissue section by immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue  underwent fixation in paraformaldehyde, heat mediated antigen retrieval in 10mM Citrate buffer pH 6.0, permeabilization in 0.1% Triton X-100 prior to blocking in 10% serum for 20 minutes at 22°C. The primary antibody, diluted 1/50 in PBS with 2% BSA and incubated with sample, for 45 minutes at 22°C. An Avidin-peroxidase conjugated horse polyclonal to rabbit Ig, was used undiluted as the secondary antibody.

    See Abreview

  • ab4052 staining Caspase-8 in formalin-fixed, paraffin-embedded Human stomach tissue by Immunohistochemistry.

References

This product has been referenced in:
  • Chang CW  et al. Lactobacillus casei Variety rhamnosus Probiotic Preventively Attenuates 5-Fluorouracil/Oxaliplatin-Induced Intestinal Injury in a Syngeneic Colorectal Cancer Model. Front Microbiol 9:983 (2018). Read more (PubMed: 29867884) »
  • Cakir U  et al. Ginger (Zingiber officinale Roscoe) for the treatment and prevention of necrotizing enterocolitis. J Ethnopharmacol 225:297-308 (2018). Read more (PubMed: 30005955) »
See all 8 Publications for this product

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

Answer

Thank you for contacting us.

I'm afraid we are unable to determine if the antibody is specific to the active form only of caspase 8 or the pro-form as it has only been tested in IHC. We also haven't tested it in staining cells (ICC/IF). Most likely it will work, as it is a simliar application to IHC, but we can't guarantee ICC/IF since it is untested.

I'm sorry I cannot be of further assistance on this occasion.

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Kidney)
Specification
Kidney
Fixative
Paraformaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM Citrate buffer, pH 6.0
Permeabilization
Yes - 0.1% Triton X-100
Blocking step
Serum as blocking agent for 20 minute(s) · Concentration: 10% · Temperature: 22°C

Dr. B Goilav

Verified customer

Submitted Dec 18 2008

Question
Answer

I'm afraid we are unable to determine if the antibody is specific to the active form only of caspase 8, as it has only been tested in IHC. I'm sorry I cannot be of further assistance on this occasion.

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Answer

I can confirm that the customer's protocol is adequate and should work. However, we have the following suggestions: 1. Try using another tissue besides liver. We recommend stomach. 2. The customer is using home brewed detection system. The sensitivity of the system may therefore not be adequate. I would suggest the followings: try using lower dilution of the primary antibody or try using more concentrated link (goat anti rabbit, 1:500 may be too diluted). Please let me know if the customer still has problems with the positive control and we can arrange for a replacement vial,

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Answer

Thank you for those further protocol details, I would like to recommend diluting the blocking serum, primary and secondary antibodies in PBST containing triton x100 (0.3%v/v) to promote permeabilization of the tissue and enable better penetration of the antibody. Please let me know if you still have problems with this antibody and we can offer you a replacement or credit note or refund if you purchased the antibody in the last 90 days,

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Answer

I'm very sorry to hear you are having problems with ab4052, thank you for taking the time to fill in the online questionnaire. Could you please tell me: -how the antibody was stored? - how long the antigen retrieval step was? - if you have done a permeabilization step? - the buffer used to dilute the antibodies? -what blocking was done (BSA/serum? HRP block?)? My apologies for the delay, thank you for your patience, I look forward to hearing from you,

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Question
Answer

Thank you for your enquiry. This antibody recognizes the full length inactive proenzyme (55 kDa). While this antibody has also been shown to detect the processed forms of Caspase 8 (which associate to form the active enzyme), its specific reaction with the active form is unknown to us. If you have any further questions then please do not hesitate to get back in touch with us.

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

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