Overview

  • Product name
  • Description
    Rabbit polyclonal to Caspase-8
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IPmore details
  • Species reactivity
    Reacts with: Mouse
    Predicted to work with: Rat, Chinese hamster
  • Immunogen

    Synthetic peptide within Mouse Caspase-8 aa 400 to the C-terminus conjugated to Keyhole Limpet Haemocyanin (KLH). The exact sequence is proprietary.
    Database link: O89110

  • Positive control
    • This antibody gave a positive signal in Raw264.7 whole cell lysate and in the following Mouse tissue lysates: Spleen; Thymus; Lung; Liver.

Properties

Applications

Our Abpromise guarantee covers the use of ab138485 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 Use a concentration of 1 µg/ml. Detects a band of approximately 58 kDa (predicted molecular weight: 55 kDa).
IP Use at 1-0.6 µg/mg of lysate.

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

  • All lanes : Anti-Caspase-8 antibody (ab138485) at 1 µg/ml (Milk blocking 3%)

    Lane 1 : Spleen (Mouse) Tissue Lysate
    Lane 2 : Thymus (Mouse) Tissue Lysate
    Lane 3 : Lung (Mouse) Whole Cell Lysate - normal tissue
    Lane 4 : Liver (Mouse) Tissue Lysate
    Lane 5 : RAW 264.7 (Mouse leukaemic monocyte macrophage cell line) Whole Cell Lysate

    Lysates/proteins at 25 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 55 kDa
    Observed band size: 58 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 24 kDa (possible non-specific binding), 36 kDa (possible non-specific binding), 48 kDa (possible non-specific binding)


    Exposure time: 8 minutes


    This blot was produced using a 10% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% Milk before being incubated with ab138485 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.

References

This product has been referenced in:
See 1 Publication for this product

Customer reviews and Q&As

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1-7 of 7 Abreviews

Application
Immunoprecipitation
Sample
Chinese hamster Cell lysate - whole cell (Chinese hamster ovary (CHO) cells)
Total protein in input
1e+006 cells
Immuno-precipitation step
Other - Protein G Dynabeads
Specification
Chinese hamster ovary (CHO) cells

Abcam user community

Verified customer

Submitted Feb 08 2017

Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (A549 (human lung epithelium cells))
Total protein in input
1e+007 cells
Immuno-precipitation step
Other - Protein G Dynabeads
Specification
A549 (human lung epithelium cells)

Abcam user community

Verified customer

Submitted Feb 08 2017

Application
Immunoprecipitation
Sample
Mouse Cell lysate - whole cell (Mouse embryonic fibroblasts (MEFs))
Total protein in input
1e+007 cells
Immuno-precipitation step
Other - Protein G Dynabeads
Specification
Mouse embryonic fibroblasts (MEFs)

Abcam user community

Verified customer

Submitted Feb 08 2017

Application
Western blot
Sample
Dog Cell lysate - whole cell (Canine Kidney Epithelial MDCK cells)
Gel Running Conditions
Reduced Denaturing (4-12% Bolt Bis-Tris Plus gel (MOPS buffer))
Loading amount
25000 cells
Specification
Canine Kidney Epithelial MDCK cells
Blocking step
Li-Cor Odyssey Blocking Buffer (TBS) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jan 17 2017

Application
Western blot
Sample
Chinese hamster Cell lysate - whole cell (Chinese hamster ovary (CHO) cells)
Gel Running Conditions
Reduced Denaturing (4-12% Bolt Bis-Tris Plus gel (MOPS buffer))
Loading amount
50000 cells
Specification
Chinese hamster ovary (CHO) cells
Blocking step
Li-Cor Odyssey Blocking Buffer (TBS) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jan 03 2017

Application
Western blot
Sample
Hamster Cell lysate - whole cell (BHK-21 (Kidney Fibroblasts))
Gel Running Conditions
Reduced Denaturing (4-12% Bolt Bis-Tris Plus gel (MOPS buffer))
Loading amount
50000 cells
Treatment
10uM Etoposide for 5 hours
Specification
BHK-21 (Kidney Fibroblasts)
Blocking step
Li-Cor Odyssey Blocking Buffer (TBS) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Dec 30 2016

Application
Immunoprecipitation
Immuno-precipitation step
Protein G
Sample
Mouse Cell lysate - whole cell (SV40-transformed MEF)
Specification
SV40-transformed MEF
Total protein in input
600 µg

Dr. Cristina Muñoz-Pinedo

Verified customer

Submitted Feb 27 2014

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