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  1. Link

    pro-caspase-8-antibody-epr162-ab108333.pdf

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Cell Biology Apoptosis Intracellular Caspases etc Caspases
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Validated using a knockout cell lineRecombinantRabMAb

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

  • Datasheet
  • SDS
Reviews (1) Submit a question References (20)

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

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR162] to Pro Caspase-8
  • Suitable for: WB, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

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Overview

  • 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. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • Jurkat, IM9, HepG2, HeLa, and HL60 cell lysates; Human tonsil tissue and Human hepatocellular carcinoma tissue; HeLa cells.
  • General notes

    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 production
    For more information see here.

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

Properties

  • 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 (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR162
  • Isotype

    IgG
  • Research areas

    • Cell Biology
    • Apoptosis
    • Intracellular
    • Caspases etc
    • Caspases
    • Cancer
    • Invasion/microenvironment
    • Apoptosis
    • Caspases
    • Cell Biology
    • Proteolysis / Ubiquitin
    • Proteolytic enzymes
    • Other proteases
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Apoptosis
    • Cancer
    • Cell Death
    • Apoptosis
    • Apoptosis Markers
    • Caspases
    • Cancer
    • Cell Death
    • Necroptosis
    • Cancer
    • Cell Death
    • Apoptosis
    • Metabolism

Associated products

  • 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

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • Jurkat cytoplasmic extract lysate (ab14842)
  • Recombinant Protein

    • Recombinant human Caspase-8 protein (ab54117)

Applications

The Abpromise guarantee

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

Antigen retrieval is recommended.

Notes
WB
1/1000 - 1/10000. Detects a band of approximately 55 kDa (predicted molecular weight: 55 kDa).
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Antigen retrieval is recommended.

Application notes
Is unsuitable for Flow Cyt or IP.

Target

  • 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

    Images

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

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

    Protocols

    • Western blot protocols
    • Immunohistochemistry protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (20)

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

    ab108333 has been referenced in 20 publications.

    • Zhang L & Su X Bioactive peptide inhibits acute myeloid leukemia cell proliferation by downregulating ALKBH5-mediated m6A demethylation of EIF4EBP1 and MLST8 mRNA. Cell Oncol (Dordr) 45:355-365 (2022). WB ; Human . PubMed: 35579750
    • Mao L  et al. Combination of oncolytic adenovirus targeting SATB1 and docetaxel for the treatment of castration-resistant prostate cancer. J Cancer 12:1846-1852 (2021). PubMed: 33613773
    • Qu L  et al. Fraxetin Inhibits the Proliferation and Metastasis of Glioma Cells by Inactivating JAK2/STAT3 Signaling. Evid Based Complement Alternat Med 2021:5540139 (2021). PubMed: 33959183
    • Wang B  et al. Combination of HDE and BIIB021 efficiently inhibits cell proliferation and induces apoptosis via downregulating hTERT in myelodysplastic syndromes. Exp Ther Med 21:503 (2021). PubMed: 33791012
    • Shi C  et al. LncRNA OIP5-AS1 promotes cell proliferation and migration and induces angiogenesis via regulating miR-3163/VEGFA in hepatocellular carcinoma. Cancer Biol Ther 21:604-614 (2020). PubMed: 32329664
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

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

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HeLa)
    Specification
    HeLa
    Fixative
    Paraformaldehyde
    Permeabilization
    Yes - 0.5% Triton-X100 in PBS
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    DR. Kirk Mcmanus

    Verified customer

    Submitted Aug 09 2012

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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