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Knockout Tested Rabbit Recombinant Monoclonal Caspase-8 antibody. Suitable for WB and reacts with Human samples. Cited in 32 publications.


Images

Western blot - Anti-Caspase-8 antibody [E7] (AB32397), expandable thumbnail
  • Western blot - Anti-Caspase-8 antibody [E7] (AB32397), expandable thumbnail
  • Western blot - Anti-Caspase-8 antibody [E7] (AB32397), expandable thumbnail
  • Western blot - Anti-Caspase-8 antibody [E7] (AB32397), expandable thumbnail
  • Western blot - Anti-Caspase-8 antibody [E7] (AB32397), expandable thumbnail

Publications

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form
Liquid
Clonality
Monoclonal

Immunogen

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

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Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
Flow CytWBICC/IF
Human
Not recommended
Tested
Not recommended

Not recommended
Not recommended

Species
Human
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
1/500 - 1/1000
Notes

For unpurified use at 1/500.

Not recommended
Not recommended

Species
Human
Dilution info
-
Notes

-

Associated Products

Select an associated product type

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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

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

Recommended products

Knockout Tested Rabbit Recombinant Monoclonal Caspase-8 antibody. Suitable for WB and reacts with Human samples. Cited in 32 publications.

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number
E7
Purification technique
Affinity purification Protein A
Specificity

The antibody should recognize both pro-form (55kDa) and p18 cleaved-form of Caspase-8.

Concentration
Loading...

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
-20°C
Aliquoting information
Upon delivery aliquot
Storage information
Avoid freeze / thaw cycle

Notes

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 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.

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.

5 product images

  • Western blot - Anti-Caspase-8 antibody [E7] (ab32397), expandable thumbnail

    Western blot - Anti-Caspase-8 antibody [E7] (ab32397)

    Lanes 1 - 4: Merged signal (red and green). Green - ab32397 observed at 55, 43/41 kDa. Red - loading control, Anti-GAPDH antibody [mAbcam 9484] - Loading Control ab9484, observed at 37 kDa.

    ab32397 was shown to specifically react with HAP1 + Staurosproin when HAP1 + Staurosproin knockout samples were used. Wild-type and HAP1 + Staurosproin knockout samples were subjected to SDS-PAGE. ab32397 and Anti-GAPDH antibody [mAbcam 9484] - Loading Control ab9484 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/500 dilution and 1/20000 dilution respectively. Blots were developed with 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 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

    All lanes: Western blot - Anti-Caspase-8 antibody [E7] (ab32397) at 1/500 dilution

    Lane 1: Wild-type HAP1 whole cell lysate at 20 µg

    Lane 2: Wild-type HAP1 whole cell lysate treated with Staurosporin at 20 µg

    Lane 3: Caspase-8 knockout HAP1 whole cell lysate at 20 µg

    Lane 4: Caspase-8 knockout HAP1 whole cell lysate treated with Staurosporin at 20 µg

    Predicted band size: 55 kDa

  • Western blot - Anti-Caspase-8 antibody [E7] (ab32397), expandable thumbnail

    Western blot - Anti-Caspase-8 antibody [E7] (ab32397)

    Lanes 1- 4: Merged signal (red and green). Green - ab32397 observed at 55 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) observed at 37 kDa.

    ab32397 was shown to react with Caspase-8 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line Human CASP8 (Caspase-8) knockout HeLa cell line ab264958 (knockout cell lysate Human CASP8 (Caspase-8) knockout HeLa cell lysate ab256857) was used. Wild-type HeLa and CASP8 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab32397 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) overnight at 4°C at a 1 in 500 dilution and a 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.

    All lanes: Western blot - Anti-Caspase-8 antibody [E7] (ab32397) at 1/500 dilution

    Lane 1: Wild-type HeLa cell lysate at 20 µg

    Lane 2: CASP8 knockout HeLa cell lysate at 20 µg

    Lane 2: Western blot - Human CASP8 (Caspase-8) knockout HeLa cell line (Human CASP8 (Caspase-8) knockout HeLa cell line ab264958)

    Lane 3: Jurkat cell lysate at 20 µg

    Lane 4: SH-SY5Y cell lysate at 20 µg

    Performed under reducing conditions.

    Predicted band size: 55 kDa

    Observed band size: 55 kDa

  • Western blot - Anti-Caspase-8 antibody [E7] (ab32397), expandable thumbnail

    Western blot - Anti-Caspase-8 antibody [E7] (ab32397)

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

    All lanes: Western blot - Anti-Caspase-8 antibody [E7] (ab32397) at 1/1000 dilution

    Lane 1: Jurkat cell lysate at 20 µg

    Lane 2: HeLa cell lysate at 20 µg

    Lane 3: IM-9 cell lysate at 20 µg

    Secondary

    All lanes: Goat Anti-Rabbit IgG, (H+L), HRP-conjugated at 1/1000 dilution

    Predicted band size: 55 kDa

  • Western blot - Anti-Caspase-8 antibody [E7] (ab32397), expandable thumbnail

    Western blot - Anti-Caspase-8 antibody [E7] (ab32397)

    All lanes: Western blot - Anti-Caspase-8 antibody [E7] (ab32397) at 1/500 dilution

    All lanes: HeLa cell lysate at 10 µg

    Predicted band size: 55 kDa

    Observed band size: 55 kDa

  • Western blot - Anti-Caspase-8 antibody [E7] (ab32397), expandable thumbnail

    Western blot - Anti-Caspase-8 antibody [E7] (ab32397)

    Western blot: Anti-CASP8 antibody [E7] (ab32397) staining at 1/500 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab32397 was shown to bind specifically to CASP8. A band was observed at 55 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in CASP8 knockout cell line. To generate this image, wild-type and CASP8 knockout HCT 116 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

    All lanes: Western blot - Anti-Caspase-8 antibody [E7] (ab32397) at 1/500 dilution

    Lane 1: Wild-type HCT 116 cell lysate at 20 µg

    Lane 2: CASP8 knockout HCT 116 cell lysate at 20 µg

    Lane 3: Wild-type HeLa cell lysate at 20 µg

    Lane 4: CASP8 knockout HeLa cell lysate at 20 µg

    Secondary

    All lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

    Performed under reducing conditions.

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Product protocols

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

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