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AB208774

Anti-Pro Caspase-8 antibody [EPR162] - BSA and Azide free

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(1 Publication)

Rabbit Recombinant Monoclonal Caspase-8 antibody. Carrier free. Suitable for WB, IHC-P and reacts with Human, Mouse, Rat samples. Cited in 1 publication.

View Alternative Names

MCH5, CASP8, Caspase-8, CASP-8, Apoptotic cysteine protease, Apoptotic protease Mch-5, CAP4, FADD-homologous ICE/ced-3-like protease, FADD-like ICE, ICE-like apoptotic protease 5, MORT1-associated ced-3 homolog, FLICE, MACH

3 Images
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Pro Caspase-8 antibody [EPR162] - BSA and Azide free (AB208774)
  • IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Pro Caspase-8 antibody [EPR162] - BSA and Azide free (AB208774)

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108333).

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

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Pro Caspase-8 antibody [EPR162] - BSA and Azide free (AB208774)
  • IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Pro Caspase-8 antibody [EPR162] - BSA and Azide free (AB208774)

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108333).

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

Western blot - Anti-Pro Caspase-8 antibody [EPR162] - BSA and Azide free (AB208774)
  • WB

Supplier Data

Western blot - Anti-Pro Caspase-8 antibody [EPR162] - BSA and Azide free (AB208774)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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] (<a href='/en-us/products/primary-antibodies/pro-caspase-8-antibody-epr162-ab108333'>ab108333</a>) at 1/1000 dilution

Lanes 3 - 4:

Western blot - Anti-Caspase-10/CASP-10 antibody [EPR10890] (<a href='/en-us/products/primary-antibodies/caspase-10-casp-10-antibody-epr10890-ab177475'>ab177475</a>) 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) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 60-70 kDa

false

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR162

Isotype

IgG

Carrier free

Yes

Reacts with

Mouse, Rat, Human

Applications

IHC-P, WB

applications

Immunogen

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

Specificity

Theoretically, this antibody should recognize p55/54, p43/41 and p18. However, in our internal tests on parent antibody ab108333, 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.

Reactivity data

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Product details

ab208774 is the carrier-free version of ab108333.

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

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

What are the advantages of a recombinant monoclonal antibody?
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.

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

Compatibility
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

Properties and storage information

Form
Liquid
Purification technique
Affinity purification Protein A
Storage buffer
pH: 7.2 - 7.4 Constituents: PBS
Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
Do Not Freeze

Supplementary information

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

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.

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 protocols

For this product, it's our understanding that no specific protocols are required. You can visit:

Target data

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). Also cleaves and activates BID, thereby promoting cytochrome C release from mitochrondria (By similarity). 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.
See full target information CASP8

Publications (1)

Recent publications for all applications. Explore the full list and refine your search

Journal of the American Academy of Dermatology 67:1327.e1-10 PubMed22884443

2012

The Fas apoptotic pathway in cutaneous T-cell lymphomas: frequent expression of phenotypes associated with resistance to apoptosis.

Applications

IHC

Species

Human

Nathalie Stutz,Ryan D Johnson,Gary S Wood
View all publications

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