Name: Anti-Pro Caspase-8 antibody [EPR162]
Brand: RabMAb
SKU: ab108333
Rating: 4 (1 reviews)

Product name

Anti-Pro Caspase-8 antibody [EPR162]
See all Pro Caspase-8 primary antibodies
Description

Rabbit monoclonal [EPR162] to Pro Caspase-8
Host species

Rabbit
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.
Tested applications

Suitable for: WB, IHC-Pmore details
Unsuitable for: Flow Cyt or IP
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Synthetic peptide within Human Pro Caspase-8 aa 350-450 (internal sequence). The exact sequence is proprietary.
Database link: Q14790
Positive control
- Jurkat, IM9, HepG2, HeLa, and HL60 cell lysates; Human tonsil tissue and Human hepatocellular carcinoma tissue; HeLa cells.
General 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 rabbit monoclonal antibody.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
Storage buffer

pH: 7.20
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 40% Glycerol, 9.85% Tris glycine, 50% Tissue culture supernatant
Purity

Tissue culture supernatant
Clonality

Monoclonal
Clone number

EPR162
Isotype

IgG
Research areas

Alternative Versions
- Anti-Pro Caspase-8 antibody [EPR162] - BSA and Azide free (ab208774)
Assay kits
- Caspase-8 Assay Kit (Colorimetric) (ab39700)
- Caspase-8 (active) FITC Staining Kit (ab65614)
Compatible Secondaries
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Positive Controls
- HepG2 cytoplasmic extract lysate (ab14659)
- Jurkat cytoplasmic extract lysate (ab14842)
Recombinant Protein
- Recombinant human Caspase-8 protein (ab54117)

Our Abpromise guarantee covers the use of ab108333 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		1/1000 - 1/10000. Detects a band of approximately 55 kDa (predicted molecular weight: 55 kDa).
IHC-P		1/100 - 1/250. Antigen retrieval is recommended.

Application notes

Is unsuitable for Flow Cyt or IP.

Relevance

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.
Database links
- Entrez Gene: 841 Human
- Entrez Gene: 12370 Mouse
- Entrez Gene: 64044 Rat
- Omim: 601763 Human
- SwissProt: Q14790 Human
- SwissProt: O89110 Mouse
- SwissProt: Q9JHX4 Rat
- Unigene: 599762 Human
- Unigene: 336851 Mouse
- Unigene: 54474 Rat
see all

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

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

Lane 1 : HepG2 whole cell lysate
Lane 2 : Rat liver tissue lysate
Lane 3 : Mouse liver tissue lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : 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

Lanes 1 - 3: Merged signal (red and green). Green - ab108333 observed at 55 kDa. Red - 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 ab8245 (loading control) overnight at 4°C. Antibody binding was detected using Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at a 1:10000 dilution for 1hr at room temperature and then imaged.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-8 antibody [EPR162] (ab108333)

ab108333, at 1/100 dilution, staining Pro Caspase-8 in paraffin-embedded human tonsil tissue by Immunohistochemistry.
Western blot - Anti-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, ab8245, observed at 37 kDa.

This western blot image is a comparison between ab108333 and a competitor's top cited mouse monoclonal antibody.
Western blot - Anti-Caspase-8 antibody [EPR162] (ab108333)

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

Lane 1 : Jurkat cell lysate
Lane 2 : IM9 cell lysate
Lane 3 : HepG2 cell lysate
Lane 4 : HL60 cell lysate

Lysates/proteins at 10 µg per lane.

Predicted band size: 55 kDa
Western blot - Anti-Caspase-8 antibody [EPR162] (ab108333)

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

Lane 1 : Jurkat cell lysate, treated with etoposide
Lane 2 : Jurkat cell lysate
Lane 3 : HeLa cell lysate, treated staurosporine
Lane 4 : HeLa cell lysate

Lysates/proteins at 10 µg per lane.

Predicted band size: 55 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-8 antibody [EPR162] (ab108333)

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

Western blot protocols
Immunohistochemistry protocols

Datasheet

This product has been referenced in:

Pittala S et al. Targeting Liver Cancer and Associated Pathologies in Mice with a Mitochondrial VDAC1-Based Peptide. Neoplasia 20:594-609 (2018). WB ; Mouse . Read more (PubMed: 29747160) »
Wang J et al. Menin mediates Tat-induced neuronal apoptosis in brain frontal cortex of SIV-infected macaques and in Tat-treated cells. Oncotarget 8:18082-18094 (2017). WB ; Rhesus monkey . Read more (PubMed: 28178646) »

See all 3 Publications for this product

Publishing research using ab108333? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-2 of 2 Abreviews or Q&A

Question

I have purchased a product from your company for my research work, I would like to know if the the product antibody can also detect the cleaved caspase -8 subunits,larger subunit p18 and smaller subunit p10, along with 55kDa . As it is imp for me to detect cleaved caspase 8 subunits for my research work. I have also read the product datasheet and it has confused me as it has not been mentioned clearly. The product datasheet reads , Specificity-A synthetic peptide corresponding to residues in human Caspase-8 was used as an immunogen. Please help me out regarding it. Product details caspase-8 rabbit monoclonal antibody clone ID: EPR162.

Abcam community

Verified customer

Asked on Aug 01 2013

Answer

The immunogen sequence is considered proprietary information. The sequence is a 30-amino acid sequence within the larger range of amino acids from 200 - 250 of the human protein sequence that is given in the UniProt entry at this link: http://www.uniprot.org/uniprot/Q14790.

Part of the sequence is in the p18 subunit and part of it is in the N-terminus propeptide. The western blot data on the datasheet suggests that the antibody is not capable of detecting the p18 subunit, or the smaller p10 subunit.

By comparison, the mouse monoclonal antibody ab119809 (clone M382) appears to be capable of detecting the full-length inactive caspase 8 and also the p10 subunit. However, we do not know of any published data for this clone.

Tom Ruyle

Abcam Scientific Support

Answered on Aug 01 2013

Application

Immunocytochemistry/ Immunofluorescence

Sample

Human Cell (HeLa)

Specification

HeLa

Fixative

Paraformaldehyde

Permeabilization

Yes - 0.5% Triton-X100 in PBS

Dr. Kirk Mcmanus

Verified customer

Submitted Aug 09 2012

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
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Recombinant Anti-Pro Caspase-8 antibody [EPR162] (ab108333)

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References

This product has been referenced in:

Customer reviews and Q&As

Immunocytochemistry/ Immunofluorescence abreview for Anti-Caspase 8 antibody [EPR162]