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Rabbit Recombinant Monoclonal Caspase-8 antibody. Suitable for WB, IHC-P and reacts with Rat, Mouse, Human samples. Cited in 34 publications.


Images

Western blot - Anti-Pro Caspase-8 antibody [EPR162] (AB108333), expandable thumbnail
  • Western blot - Anti-Pro Caspase-8 antibody [EPR162] (AB108333), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Pro Caspase-8 antibody [EPR162] (AB108333), expandable thumbnail
  • Western blot - Anti-Pro Caspase-8 antibody [EPR162] (AB108333), expandable thumbnail
  • Western blot - Anti-Pro Caspase-8 antibody [EPR162] (AB108333), expandable thumbnail

Publications

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.2 - 7.4
Preservative: 0.05% Sodium azide
Constituents: 50% Tissue culture supernatant, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 0.1% BSA

Form
Liquid
Clonality
Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
WBIHC-PFlow CytIP
Human
Tested
Tested
Not recommended
Not recommended
Mouse
Tested
Expected
Not recommended
Not recommended
Rat
Tested
Expected
Not recommended
Not recommended

Tested
Tested

Species
Rat
Dilution info
1/1000 - 1/10000
Notes

-

Species
Mouse
Dilution info
1/1000 - 1/10000
Notes

-

Species
Human
Dilution info
1/1000 - 1/10000
Notes

-

Tested
Tested

Species
Human
Dilution info
1/100 - 1/250
Notes

Antigen retrieval is recommended.

Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Expected
Expected

Species
Rat, Mouse
Dilution info
Use at an assay dependent concentration.
Notes

-

Not recommended
Not recommended

Species
Human, Mouse, Rat
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Human, Mouse, Rat
Dilution info
-
Notes

-

Associated Products

Select an associated product type

10 products for Alternative Product

Target data

Function

Thiol protease that plays a key role in programmed cell death by acting as a molecular switch for apoptosis, necroptosis and pyroptosis, and is required to prevent tissue damage during embryonic development and adulthood (PubMed:23516580, PubMed:35338844, PubMed:35446120, PubMed:8681376, PubMed:8681377, PubMed:8962078, PubMed:9006941, PubMed:9184224). Initiator protease that induces extrinsic apoptosis by mediating cleavage and activation of effector caspases responsible for FAS/CD95-mediated and TNFRSF1A-induced cell death (PubMed:23516580, PubMed:35338844, PubMed:35446120, PubMed:8681376, PubMed:8681377, PubMed:8962078, PubMed:9006941, PubMed:9184224). Cleaves and activates effector caspases CASP3, CASP4, CASP6, CASP7, CASP9 and CASP10 (PubMed:16916640, PubMed:8962078, PubMed:9006941). Binding to the adapter molecule FADD recruits it to either receptor FAS/TNFRSF6 or TNFRSF1A (PubMed:8681376, PubMed:8681377). The resulting aggregate called the death-inducing signaling complex (DISC) performs CASP8 proteolytic activation (PubMed:9184224). The active dimeric enzyme is then liberated from the DISC and free to activate downstream apoptotic proteases (PubMed:9184224). Proteolytic fragments of the N-terminal propeptide (termed CAP3, CAP5 and CAP6) are likely retained in the DISC (PubMed:9184224). In addition to extrinsic apoptosis, also acts as a negative regulator of necroptosis: acts by cleaving RIPK1 at 'Asp-324', which is crucial to inhibit RIPK1 kinase activity, limiting TNF-induced apoptosis, necroptosis and inflammatory response (PubMed:31827280, PubMed:31827281). Also able to initiate pyroptosis by mediating cleavage and activation of gasdermin-C and -D (GSDMC and GSDMD, respectively): gasdermin cleavage promotes release of the N-terminal moiety that binds to membranes and forms pores, triggering pyroptosis (PubMed:32929201, PubMed:34012073). Initiates pyroptosis following inactivation of MAP3K7/TAK1 (By similarity). Also acts as a regulator of innate immunity by mediating cleavage and inactivation of N4BP1 downstream of TLR3 or TLR4, thereby promoting cytokine production (By similarity). May participate in the Granzyme B (GZMB) cell death pathways (PubMed:8755496). Cleaves PARP1 and PARP2 (PubMed:8681376). Independent of its protease activity, promotes cell migration following phosphorylation at Tyr-380 (PubMed:18216014, PubMed:27109099). Isoform 5. Lacks the catalytic site and may interfere with the pro-apoptotic activity of the complex. Isoform 6. Lacks the catalytic site and may interfere with the pro-apoptotic activity of the complex. Isoform 7. Lacks the catalytic site and may interfere with the pro-apoptotic activity of the complex (Probable). Acts as an inhibitor of the caspase cascade (PubMed:12010809). Isoform 8. Lacks the catalytic site and may interfere with the pro-apoptotic activity of the complex.

Alternative names

Recommended products

Rabbit Recombinant Monoclonal Caspase-8 antibody. Suitable for WB, IHC-P and reacts with Rat, Mouse, Human samples. Cited in 34 publications.

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number
EPR162
Purification technique
Affinity purification Protein A
Specificity

Theoretically, ab108333 should recognize p55/54, p43/41 and p18. However, in our internal tests, it only recognized pro caspase 8. We tested it side by side with ab32397. Both antibodies are KO validated, and ab32397 recognizes pro-caspase 8 and cleavage caspase 8.

Concentration
Loading...

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
-20°C
Storage information
Stable for 12 months at -20°C

Notes

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 monoclonal antibody, which offers several advantages including:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

Pro Caspase-8 also known as CASP8 is an inactive precursor of caspase-8 an important enzyme in the apoptosis and necroptosis pathways. This protein has a molecular mass of approximately 55 kDa. Caspase-8 expression occurs in various tissues with higher prevalence in immune cells including T lymphocytes and natural killer cells. Researchers often use techniques like western blot for caspase 8 detection particularly when analyzing its activation state.

Biological function summary

Pro caspase-8 plays a significant role in the regulation of apoptosis a programmed cell death process. It is initially synthesized in an inactive form and requires activation by cleavage to become functional. Pro caspase-8 integrates into the death-inducing signaling complex (DISC) following specific triggers such as Fas receptor engagement or TNF receptor activation indicating its role in response to external apoptotic signals. Its activation marks the initiation of a cascade that results in apoptosis ensuring controlled cell removal in multicellular organisms.

Pathways

Pro caspase-8 functions within the extrinsic apoptosis pathway where it serves as an initiator caspase. Alongside other proteins like Fas-associated protein with death domain (FADD) caspase-8 forms a critical part of the apoptotic signaling network. Another important pathway involving pro caspase-8 is necroptosis an alternative form of programmed cell death where it can inhibit the function of receptor-interacting protein kinase 1 (RIPK1) and RIPK3 under certain conditions. The regulation of these pathways highlights pro caspase-8's central role in determining cell fate under stress or damage.

Associated diseases and disorders

Aberrations in pro caspase-8 activity link to various conditions such as cancer and autoimmune diseases. In cancers insufficient caspase-8 activation results in the evasion of apoptosis allowing uncontrolled cell proliferation. Meanwhile excessive or improper activation associates with autoimmune disorders where unregulated apoptosis leads to immune system imbalance. Pro caspase-8 connects with proteins like FLICE-like inhibitory protein (c-FLIP) in cancer where it influences the apoptotic threshold and contributes to the pathology.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

8 product images

  • Western blot - Anti-Pro Caspase-8 antibody [EPR162] (ab108333), expandable thumbnail

    Western blot - Anti-Pro Caspase-8 antibody [EPR162] (ab108333)

    Lane 1: Wild-type HAP1 cell lysate (20 μg)
    Lane 2: Pro Caspase-8 knockout HAP1 cell lysate (20 μg)
    Lanes 1 and 2: Merged signal (red and green). Green - ab108333 observed at 55 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245, observed at 37 kDa.

    ab108333 was shown to specifically react with Caspase-8 when Caspase-8 knockout samples were used. Wild-type and Pro Caspase-8 knockout samples were subjected to SDS-PAGE. ab108333 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (loading control to GAPDH) were both diluted 1/1000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (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.

    All lanes: Western blot - Anti-Pro Caspase-8 antibody [EPR162] (ab108333)

    Predicted band size: 55 kDa

  • Western blot - Anti-Pro Caspase-8 antibody [EPR162] (ab108333), expandable thumbnail

    Western blot - Anti-Pro Caspase-8 antibody [EPR162] (ab108333)

    Lanes 1 - 3: Merged signal (red and green). Green - ab108333 observed at 55 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245, observed at 37 kDa.

    This blot was produced using a 4-12% 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 Licor blocking buffer before being incubated with ab108333 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (loading control) overnight at 4°C. Antibody binding was detected using Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at a 1:10000 dilution for 1hr at room temperature and then imaged.

    All lanes: Western blot - Anti-Pro Caspase-8 antibody [EPR162] (ab108333) at 1/1000 dilution

    Lane 1: HepG2 whole cell lysate at 20 µg

    Lane 2: Rat liver tissue lysate at 20 µg

    Lane 3: Mouse liver tissue lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size: 55 kDa

    Observed band size: 55 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Pro Caspase-8 antibody [EPR162] (ab108333), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Pro Caspase-8 antibody [EPR162] (ab108333)

    ab108333, at 1/100 dilution, staining Pro Caspase-8 in paraffin-embedded human tonsil tissue by Immunohistochemistry.

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Western blot - Anti-Pro Caspase-8 antibody [EPR162] (ab108333), expandable thumbnail

    Western blot - Anti-Pro Caspase-8 antibody [EPR162] (ab108333)

    Lane 1: Wild-type HAP1 cell lysate (20 μg)
    Lane 2: Pro Caspase-8 knockout HAP1 cell lysate (20 μg)
    Lanes 1 and 2: Merged signal (red and green).

    Green - target observed at 55 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245, observed at 37 kDa.

    This western blot image is a comparison between ab108333 and a competitor's top cited mouse monoclonal antibody.

    All lanes: Western blot - Anti-Pro Caspase-8 antibody [EPR162] (ab108333)

    Predicted band size: 55 kDa

  • Western blot - Anti-Pro Caspase-8 antibody [EPR162] (ab108333), expandable thumbnail

    Western blot - Anti-Pro Caspase-8 antibody [EPR162] (ab108333)

    All lanes: Western blot - Anti-Pro Caspase-8 antibody [EPR162] (ab108333) at 1/1000 dilution

    Lane 1: Jurkat cell lysate at 10 µg

    Lane 2: IM9 cell lysate at 10 µg

    Lane 3: HepG2 cell lysate at 10 µg

    Lane 4: HL60 cell lysate at 10 µg

    Predicted band size: 55 kDa

  • Western blot - Anti-Pro Caspase-8 antibody [EPR162] (ab108333), expandable thumbnail

    Western blot - Anti-Pro Caspase-8 antibody [EPR162] (ab108333)

    All lanes: Western blot - Anti-Pro Caspase-8 antibody [EPR162] (ab108333) at 1/1000 dilution

    Lane 1: Jurkat cell lysate, treated with etoposide at 10 µg

    Lane 2: Jurkat cell lysate at 10 µg

    Lane 3: HeLa cell lysate, treated staurosporine at 10 µg

    Lane 4: HeLa cell lysate at 10 µg

    Predicted band size: 55 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Pro Caspase-8 antibody [EPR162] (ab108333), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Pro Caspase-8 antibody [EPR162] (ab108333)

    ab108333, at 1/100 dilution, staining Pro Caspase-8 in paraffin-embedded human hepatocellular carcinoma by Immunohistochemistry.

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Western blot - Anti-Pro Caspase-8 antibody [EPR162] (ab108333), expandable thumbnail

    Western blot - Anti-Pro Caspase-8 antibody [EPR162] (ab108333)

    Blocking and diluting buffer: 5% NFDM /TBST.

    Exposure time:

    Lane 1 & 2: 7 seconds

    Lane 3 & 4: 3 seconds

    Lanes 1 - 2: Western blot - Anti-Pro Caspase-8 antibody [EPR162] (ab108333) at 1/1000 dilution

    Lanes 3 - 4: Western blot - Anti-Caspase-10/CASP-10 antibody [EPR10890] (Anti-Caspase-10/CASP-10 antibody [EPR10890] ab177475) at 1/1000 dilution

    Lanes 1 - 2: His-tagged Human full-length Caspase 8 recombinant protein at 10 ng

    Lanes 3 - 4: His-tagged Human full-length Caspase 10 recombinant protein at 10 ng

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Observed band size: 60-70 kDa

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