Validated using a knockout cell line
Recombinant
RabMAb

Anti-Caspase-8 antibody [E6] (ab32125)

Overview

  • Product name
    Anti-Caspase-8 antibody [E6]
    See all Caspase-8 primary antibodies
  • Description
    Rabbit monoclonal [E6] to Caspase-8
  • Specificity
    ab32125 should recognize all splice isoforms of Caspase-8.
  • Tested applications
    Suitable for: WBmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Caspase-8 aa 1-100 (N terminal).
    Database link: Q14790

  • Epitope
    ab32125 reacts with an epitope located in the N terminal region of caspase-8.
  • Positive control
    • Jurkat cell lysate, human breast adenocarcinoma.
  • General notes

     

    A trial size is available to purchase for this antibody.

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab32125 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
WB 1/3000. Detects a band of approximately 55 kDa (predicted molecular weight: 55 kDa).

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

  • Anti-Caspase-8 antibody [E6] (ab32125) at 1/3000 dilution + Jurkat cell lysate

    Predicted band size : 55 kDa
    Observed band size : 55 kDa


  • Predicted band size : 55 kDa

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: Caspase-8 knockout HAP1 cell lysate (20 µg)
    Lanes 1 and 2: Merged signal (red and green). Green - ab32125 observed at 55 kDa. Red - loading control, ab8226, observed at 42 kDa.
    ab32125 was shown to recognize Caspase-8 when Caspase-8 knockout samples were used, along with additional cross-reactive bands. Wild-type and Caspase-8 knockout samples were subjected to SDS-PAGE. ab32125 and ab8226 (loading control to beta actin) were diluted 1/3000 and 1/1000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

References

This product has been referenced in:
  • Iwasaki K  et al. Cytotoxic effects of the radiocontrast agent iotrolan and anesthetic agents bupivacaine and lidocaine in three-dimensional cultures of human intervertebral disc nucleus pulposus cells: identification of the apoptotic pathways. PLoS One 9:e92442 (2014). WB ; Human . Read more (PubMed: 24642945) »
  • Huet HA  et al. Multivalent nanobodies targeting death receptor 5 elicit superior tumor cell killing through efficient caspase induction. MAbs 6:1560-70 (2014). IP . Read more (PubMed: 25484045) »

See all 10 Publications for this product

Customer reviews and Q&As

Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing
Sample
Mouse Cell lysate - whole cell (NMuMG cells)
Specification
NMuMG cells
Treatment
TGF-beta, 24 hours, 50 pM
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C
Username

Abcam user community

Verified customer

Submitted May 07 2014

Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing
Sample
Human Cell lysate - whole cell (RAMOS cells)
Specification
RAMOS cells
Treatment
TGF-beta, 24 hours, 50 pM
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C
Username

Abcam user community

Verified customer

Submitted May 07 2014

Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing
Sample
African Green Monkey Cell lysate - whole cell (COS-1 cells)
Specification
COS-1 cells
Treatment
TGF-beta, 24 hours, 50 pM
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C
Username

Abcam user community

Verified customer

Submitted May 07 2014

Thank you for your inquiry.
I am happy to confirm that all these antibodies are tested and guaranteed to work with human samples.
You are correct that we do not have any reports yet about the use of these antibodies in muscle. I assume that i...

Read More

Thank you for your enquiry. I can confirm that the peptide sequence : DFSRNLYDIGEQLDSEDLAC I hope this information will be useful to you. Should you require any other information, please do not hesitate to contact me.

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

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