Anti-Caspase-8 抗体 (ab4052)

Reviews (1)Q&A (6)References (10)

Key features and details

  • Rabbit polyclonal to Caspase-8
  • Suitable for: IHC-P
  • Reacts with: Mouse, Rat, Cow, Human, Monkey
  • Isotype: IgG

製品の概要

  • 製品名

    Anti-Caspase-8 antibody
    Caspase-8 一次抗体 製品一覧

  • 製品の詳細

    Rabbit polyclonal to Caspase-8

  • 由来種

    Rabbit

  • アプリケーション

    適用あり: IHC-Pmore details

  • 種交差性

    交差種: Mouse, Rat, Cow, Human, Monkey

  • 免疫原

    Recombinant full length protein corresponding to Human Caspase-8.
    Database link: Q14790

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab4052 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
IHC-P 1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

ターゲット情報

  • 機能

    Most upstream protease of the activation cascade of caspases responsible for the TNFRSF6/FAS mediated and TNFRSF1A induced cell death. Binding to the adapter molecule FADD recruits it to either receptor. The resulting aggregate called death-inducing signaling complex (DISC) performs CASP8 proteolytic activation. The active dimeric enzyme is then liberated from the DISC and free to activate downstream apoptotic proteases. Proteolytic fragments of the N-terminal propeptide (termed CAP3, CAP5 and CAP6) are likely retained in the DISC. Cleaves and activates CASP3, CASP4, CASP6, CASP7, CASP9 and CASP10. May participate in the GZMB apoptotic pathways. Cleaves ADPRT. Hydrolyzes the small-molecule substrate, Ac-Asp-Glu-Val-Asp-
    -AMC. Likely target for the cowpox virus CRMA death inhibitory protein. Isoform 5, isoform 6, isoform 7 and isoform 8 lack the catalytic site and may interfere with the pro-apoptotic activity of the complex.

  • 組織特異性

    Isoform 1, isoform 5 and isoform 7 are expressed in a wide variety of tissues. Highest expression in peripheral blood leukocytes, spleen, thymus and liver. Barely detectable in brain, testis and skeletal muscle.

  • 関連疾患

    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.

  • 配列類似性

    Belongs to the peptidase C14A family.
    Contains 2 DED (death effector) domains.

  • ドメイン

    Isoform 9 contains a N-terminal extension that is required for interaction with the BCAP31 complex.

  • 翻訳後修飾

    Generation of the subunits requires association with the death-inducing signaling complex (DISC), whereas additional processing is likely due to the autocatalytic activity of the activated protease. GZMB and CASP10 can be involved in these processing events.
    Phosphorylated upon DNA damage, probably by ATM or ATR.

  • 細胞内局在

    Cytoplasm.
  • Information by UniProt

  • 参照データベース

    see all

  • 別名

    • ALPS2B antibody
    • Amyotrophic lateral sclerosis 2 chromosomal region candidate gene 12 protein antibody
    • Apoptosis related cysteine peptidase antibody
    • Apoptotic cysteine protease antibody
    • Apoptotic protease Mch-5 antibody
    • Apoptotic protease Mch5 antibody
    • CAP 4 antibody
    • CAP4 antibody
    • CASP-8 antibody
    • CASP8 antibody
    • CASP8_HUMAN antibody
    • Caspase 8 antibody
    • Caspase 8 apoptosis related cysteine peptidase antibody
    • Caspase IIX antibody
    • Caspase-8 subunit p10 antibody
    • caspase8 antibody
    • CED 3 antibody
    • FADD Homologous ICE/CED3 Like Protease antibody
    • FADD Like ICE antibody
    • FADD-homologous ICE/CED-3-like protease antibody
    • FADD-like ICE antibody
    • FLICE antibody
    • FLJ17672 antibody
    • ICE-like apoptotic protease 5 antibody
    • MACH alpha 1/2/3 protein antibody
    • MACH antibody
    • MACH beta 1/2/3/4 protein antibody
    • MACH5 antibody
    • MCH 5 antibody
    • MCH5 antibody
    • MGC78473 antibody
    • MORT1 associated ced 3 homolog antibody
    • MORT1 associated CED3 homolog antibody
    • MORT1-associated CED-3 homolog antibody
    • OTTHUMP00000163717 antibody
    • OTTHUMP00000163720 antibody
    • OTTHUMP00000163724 antibody
    • OTTHUMP00000163725 antibody
    • OTTHUMP00000165062 antibody
    • OTTHUMP00000165063 antibody
    • OTTHUMP00000165064 antibody
    • OTTHUMP00000206552 antibody
    • OTTHUMP00000206582 antibody
    see all

画像

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-8 antibody (ab4052)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-8 antibody (ab4052)This image is courtesy of an Abreview submitted by Dr B Goilav

    ab4052 staining Caspase-8 in human kidney tissue section by immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue  underwent fixation in paraformaldehyde, heat mediated antigen retrieval in 10mM Citrate buffer pH 6.0, permeabilization in 0.1% Triton X-100 prior to blocking in 10% serum for 20 minutes at 22°C. The primary antibody, diluted 1/50 in PBS with 2% BSA and incubated with sample, for 45 minutes at 22°C. An Avidin-peroxidase conjugated horse polyclonal to rabbit Ig, was used undiluted as the secondary antibody.

    See Abreview

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-8 antibody (ab4052)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-8 antibody (ab4052)

    ab4052 staining Caspase-8 in formalin-fixed, paraffin-embedded Human stomach tissue by Immunohistochemistry.

参考文献 (10)

ab4052 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab4052 は 10 報の論文で使用されています。

  • Buyuktiryaki M  et al. Therapeutic and preventative effects of ankaferd blood stopper in an experimental necrotizing enterocolitis model. Biomed Pharmacother 110:105-110 (2019). PubMed: 30465999
  • Chang CW  et al. Lactobacillus casei Variety rhamnosus Probiotic Preventively Attenuates 5-Fluorouracil/Oxaliplatin-Induced Intestinal Injury in a Syngeneic Colorectal Cancer Model. Front Microbiol 9:983 (2018). PubMed: 29867884
  • Cakir U  et al. Ginger (Zingiber officinale Roscoe) for the treatment and prevention of necrotizing enterocolitis. J Ethnopharmacol 225:297-308 (2018). PubMed: 30005955
  • Masalova OV  et al. Modulation of Cell Death Pathways by Hepatitis C Virus Proteins in Huh7.5 Hepatoma Cells. Int J Mol Sci 18:N/A (2017). PubMed: 29113144
  • Yao Q  et al. Prognostic significance of TRAIL signalling molecules in cervical squamous cell carcinoma. J Clin Pathol 69:122-7 (2016). PubMed: 26254281
  • Molteno AC  et al. Otago Glaucoma Surgery Outcome Study: Further Histology and Immunohistochemistry of Molteno Implant Blebs. Invest Ophthalmol Vis Sci 56:4364-74 (2015). PubMed: 26176873
  • Negrette-Guzmán M  et al. Sulforaphane attenuates gentamicin-induced nephrotoxicity: role of mitochondrial protection. Evid Based Complement Alternat Med 2013:135314 (2013). PubMed: 23662110
  • Sansanwal P  et al. Caspase-4 may play a role in loss of proximal tubules and renal injury in nephropathic cystinosis. Pediatr Nephrol : (2009). IHC-P ; Human . PubMed: 19705160
  • Wu LP  et al. HDAC inhibitor depsipeptide activates silenced genes through decreasing both CpG and H3K9 methylation on the promoter. Mol Cell Biol : (2008). PubMed: 18332107
  • Goilav B  et al. Pathways of apoptosis in human autosomal recessive and autosomal dominant polycystic kidney diseases. Pediatr Nephrol 23:1473-82 (2008). IHC-P ; Human . PubMed: 18516626

レビューと Q&A

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1-7 of 7 Abreviews or Q&A

Question

I am not clear that whether ab4052(anti-caspase 8) will be recognizing full length caspase 8(pro-caspase 8). Does this antibody work for cell culture staining (IFC)? Thanks

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Answer

Thank you for contacting us.

I'm afraid we are unable to determine if the antibody is specific to the active form only of caspase 8 or the pro-form as it has only been tested in IHC. We also haven't tested it in staining cells (ICC/IF). Most likely it will work, as it is a simliar application to IHC, but we can't guarantee ICC/IF since it is untested.

I'm sorry I cannot be of further assistance on this occasion.

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Caspase 8 antibody

 Excellent
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Kidney)
Specification
Kidney
Fixative
Paraformaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM Citrate buffer, pH 6.0
Permeabilization
Yes - 0.1% Triton X-100
Blocking step
Serum as blocking agent for 20 minute(s) · Concentration: 10% · Temperature: 22°C
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Dr. B Goilav

Verified customer

投稿 Dec 18 2008

Question

Does the antibody recognise the active form only of caspase 8?

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Answer

I'm afraid we are unable to determine if the antibody is specific to the active form only of caspase 8, as it has only been tested in IHC. I'm sorry I cannot be of further assistance on this occasion.

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Question

Christian is visting Cambridge (and Abcam of course), so I forward our customers reply to your suggestion: "... Dear abcam-team Thank you for your kind reply concerning the antibody anti-Caspase 8 / Flice no. AB4052. I have used the PBST medium as diluent for the antibodies, but again failed to stain apoptotic cells. Further changes in our protocol were also not successful. I got no staining at all in any slide. You successfully used this antibody in immunohistochemistry in paraffin embedded murine tissue. Would it be possible to get your staining-protocol? It would help me very much. Thanks in advance J. Podlech .." Hoepfully you can provide such a protocol?

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Answer

I can confirm that the customer's protocol is adequate and should work. However, we have the following suggestions: 1. Try using another tissue besides liver. We recommend stomach. 2. The customer is using home brewed detection system. The sensitivity of the system may therefore not be adequate. I would suggest the followings: try using lower dilution of the primary antibody or try using more concentrated link (goat anti rabbit, 1:500 may be too diluted). Please let me know if the customer still has problems with the positive control and we can arrange for a replacement vial,

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Question

follow up to previous correspondance: The antibody was stored in our refrigerator (4°C) and used the other day, after delivery. The antigen retrieval step was performed as we usually do it. That means a boiling step of about 5 min and cooling for about 1 hour in the unmasking solution. It works very well with a lot of antibodies in our hands. A permeabilisation step was not done, because I don't think that this step is really necessary for formalin-fixed and paraffin-embedded tissue. The antibody was diluted in TBS. The immunohistochemistry protocol includes a serum block (10% goat serum in TBS) before incubation with the first antibody. I hope this further information is helpful. Thank you very much for your efforts.

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Answer

Thank you for those further protocol details, I would like to recommend diluting the blocking serum, primary and secondary antibodies in PBST containing triton x100 (0.3%v/v) to promote permeabilization of the tissue and enable better penetration of the antibody. Please let me know if you still have problems with this antibody and we can offer you a replacement or credit note or refund if you purchased the antibody in the last 90 days,

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Question

BATCH NUMBER 98717 ORDER NUMBER -- NOT SPECIFIED -- DESCRIPTION OF THE PROBLEM This antibody does not work in immunohistochemistry with murine liver tissue fixes with 4% buffered formalin and embedded in paraffin with routine methods in our hands. We got no staining. SAMPLE Mouse Liver (whole organ) SECONDARY ANTIBODY anti-Rabbit conjugated with biotin, 1:500, from Sigma B8895, for 30 min at RT DETECTION METHOD the tissue used is positive with an anti-active Caspase 3 ab and serves generally for me as an apoptosis positive control with a lot of microscopically typical apoptotic cells (FAS induced) POSITIVE AND NEGATIVE CONTROLS USED FAS induced liver cell necroses FIXATION OF SAMPLE Formalin 4%, buffered with PBS to pH=7.4 ANTIGEN RETRIEVAL anti-Caspase 8, 0.5 mL vial, used in a 1:50 dilution in TBS for O/N at room temperature BLOCKING CONDITIONS enzymatic digestion with trypsin or without digestion, followed by the unmasking technique with citrate buffer before incubation with the primary antibody. Detection with the Avidin-Biotin-Complex-method (peroxidase) and DAB as the substrate after incubation with the secondary antibody. HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 4 HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes ADDITIONAL NOTES Other anti caspase antibody work well in this system

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Answer

I'm very sorry to hear you are having problems with ab4052, thank you for taking the time to fill in the online questionnaire. Could you please tell me: -how the antibody was stored? - how long the antigen retrieval step was? - if you have done a permeabilization step? - the buffer used to dilute the antibodies? -what blocking was done (BSA/serum? HRP block?)? My apologies for the delay, thank you for your patience, I look forward to hearing from you,

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Question

Will this product detect both the active an inactive form of caspase 8?

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Answer

Thank you for your enquiry. This antibody recognizes the full length inactive proenzyme (55 kDa). While this antibody has also been shown to detect the processed forms of Caspase 8 (which associate to form the active enzyme), its specific reaction with the active form is unknown to us. If you have any further questions then please do not hesitate to get back in touch with us.

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