Caspase-8 Antibody (90A992)

Western Blot: Caspase-8 Antibody (90A992) [NB100-56527] - Western blot analysis of Caspase-8 in Jurkat cells using Caspase-8 antibody at 1 ug/ml. Cells were treated with 2 uM staurosporine for different time periods. Caspase-8 activation is detected in western blots by the presence of Caspase-8 cleavage fragments. The antibody detected both pro (full length) and active (cleaved) Caspase-8, depending on the treatment time points. A basal level of endogenously cleaved Caspase-8 can be see in untreated Jurkat cells. Goat anti-mouse lg HRP secondary antibody and PicoTect ECL substrate solution were used for this test.

Western Blot: Caspase-8 Antibody (90A992) [NB100-56527] - Caspase-8 expression in 786-O and A-498 cellsClose

Immunohistochemistry-Paraffin: Caspase-8 Antibody (90A992) [NB100-56527] - Formalin-fixed, paraffin-embedded human spleen (top) and breast (bottom) stained with Caspase-8 antibody at 4 ug/ml. Localization can be cytoplasmic and nuclear. Cancer/normal adjacent tissue array was used for this test. Staining of formalin-fixed tissues is enhanced by boiling tissue sections in 10 mM sodium citrate buffer, pH 6.0 for 10-20 min followed by cooling at RT for 20 min.

Flow Cytometry: Caspase-8 Antibody (90A992) [NB100-56527] - Flow cytometric analysis of Caspase-8 in HeLa cells using 0.1 ug of Caspase-8 antibody. Shaded histogram represents cells without antibody; green represents isotype control; red represents Caspase-8 antibody. Goat anti-mouse IgG-FITC secondary antibody was used for this test. IC-Flow (Intracellular Staining Flow Cytometry Kit) was used to fix and prepare the cells for staining.

Western Blot: Caspase-8 Antibody (90A992) [NB100-56527] - Analysis using the Biotin conjugate of NB100-56527. Detection of human Caspase-8 using Jurkat lysates with NB100-55786 at 2 ug/ml (lane 1) and 0.5 ug/ml (lane 2) dilution. NB100-55786 only detects 55 kDa Caspase-8 in Jurkat cells.

Simple Western: Caspase-8 Antibody (90A992) [NB100-56527] - Simple Western lane view shows a specific band for Caspase 8 in 0.5 mg/ml of Hek293 lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

• Reactivity: Hu, Pm
• Applications: WB, Simple Western, Flow, IHC, IHC-P, CyTOF-ready
• Clone: 90A992
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: 1.0 mg/ml

Caspase-8 Antibody (90A992) Summary

• Immunogen: The immunogen for this product falls between amino acids 360-385 of the human Caspase 8 protein.
• Specificity: This can be used for detection of the pro-form of Caspase-8.
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Gene: CASP8
• Purity: Protein G purified

Applications/Dilutions

• Dilutions:
  • Western Blot 0.5-2 ug/ml
  • Simple Western 1:100
  • Flow Cytometry 0.1-0.5 ug/ml
  • Immunohistochemistry
  • Immunohistochemistry-Paraffin 4 ug/ml
  • CyTOF-ready
• Application Notes: In Simple Western only 10 - 15 ul of the recommended dilution is used per data point. Separated by Size. This antibody is CyTOF ready. Use in Immunohistochemistry reported in scientific literature (PMID: 31034097).
• Control:

  Jurkat Whole Cell Lysate
  Caspase-8 Overexpression Lysate
  Caspase-8 Knockout HeLa Cell Lysate
  Caspase-8 Knockout HeLa Cell Lysate

Reactivity Notes

Chimpanzee and Rhesus Monkey reactivity noted.

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.05% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Protein G purified

Alternate Names for Caspase-8 Antibody (90A992)

• AIS
• androgen receptor
• CASP8
• Caspase8
• Caspase-8
• DHTRTFM
• Dihydrotestosterone receptorHYSP1
• HUMARA
• Mch5
• NR3C4KD
• Nuclear receptor subfamily 3 group C member 4
• SMAX1SBMA
• spinal and bulbar muscular atrophy

Background

Caspases are a family of cysteine proteases that are key mediators of programmed cell death or apoptosis. The precursor form of all caspases is composed of a prodomain, and large and small catalytic subunits. The active forms of caspases are generated by several stimuli including ligand-receptor interactions, growth factor deprivation and inhibitors of cellular functions. All known caspases require cleavage adjacent to aspartates to liberate one large and one small subunit, which associate into a2b2 tetramer to form the active enzyme. Caspase-8 (FLICE) forms a direct link between the activation of CD95 and the caspase pathway. Overexpression of Caspase-8 induces apoptosis, which can be blocked by inhibitors specific for the ICE family.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Caspase-8 Antibody (NB100-56527)(8)

Publications using NB100-56527

• Wu SH, Chyau CC, Chen JH et al. Anti-cancerous effects of Wasabia japonica extract in Hep3B liver cancer cells via ROS accumulation, DNA damage and p73-mediated apoptosis Journal of Functional Foods 2015 Mar 02 (WB, Human)
• Toy R, Pradhan P, Ramesh V et al. Mutational landscape of penile squamous cell carcinoma in a Chinese population Int. J. Cancer Apr 29 2019 [PMID: 31034097] (IHC)
• Greenberg DE, Sturdevant DE, Marshall-Batty KR et al. Simultaneous Host-Pathogen transcriptome analysis during Granulibacter bethesdensis infection of normal and chronic granulomatous disease neutrophils. Infect. Immun. 2015 Aug 17 [PMID: 26283340] (WB, Human)
• Hsuan SW, Chyau CC, Hung HY et al. The induction of apoptosis and autophagy by Wasabia japonica extract in colon cancer Eur J Nutr 2015 Feb 27 [PMID: 25720497] (WB, Human)
• Cheng Tc, Lai Cs, Chung Mc et al. Potent anti-cancer effect of 3'-hydroxypterostilbene in human colon xenograft tumors PLoS OnE et al. 2014 Nov 13 [PMID: 25389774] (WB, Human)
  
  Details:
  Caspase-8 antibody used in WB for the detection of pro and cleaved forms of Caspase 8 protein in lysates of COLO205 cancer cells treated or not with Pterostilbene and 3'-hydroxypterostilbene (Figure 2C)
• Schultz CR, Golembieski WA, King DA et al. Inhibition of HSP27 alone or in combination with pAKT inhibition as therapeutic approaches to target SPARC-induced glioma cell survival. Mol Cancer. 2012 Apr 5 [PMID: 22480225]
• Berges C, Haberstock H, Fuchs D et al. Proteasome inhibition activates the mitochondrial pathway of apoptosis in human CD4+ T cells. J Cell Biochem. 2009 Nov 1 [PMID: 19735079] (WB)
  
  Details:
  Caspase-8 Pro and Active (IMG-274A). 1.WB: Purified CD4+ T cells treated with DMSO or bortezomib (Bor), Fig 1C. 2. WB: CD4+ T cells pretreated with ZVAD followed by Bor or DMSO incubation, Fig 3.
• Berges C, Fuchs D, Opelz G et al. Helenalin suppresses essential immune functions of activated CD4+ T cells by multiple mechanisms. Mol Immunol. 2009 Sep [PMID: 19656571] (WB)
  
  Details:
  Caspase-8 Pro and Active (IMG-274A). WB: Activated CD4+ T cells were treated with helenalin or DMSO, Fig 2. Note: Caspase 8 was seen at 55 kDa and a doublet at 43 kDa, Fig 2A.

Review for Caspase-8 Antibody (NB100-56527) (1)

Average Rating: 4

(Based on 1 review)

We have 1 review tested in 1 species: Human.

reviewed by: Anonymous WB Human 04/29/2015

Summary

• Application: Western Blot
• Sample Tested: 786-O and A-498 whole cell lysates
• Species: Human

Blocking

• Blocking Details: n/a

Details

• Detection Notes: ECL, UVP Biospectrum TM500 imaging system, exposure time p43/p41: 5-10 min; p18: 30min

FAQs for Caspase-8 Antibody (NB100-56527). (Showing 1 - 2 of 2 FAQs).

1. May we ask the MW difference between the active form and the proform of Caspase 8 while performing WB?
   • In treated cells induced to undergo apoptosis, caspase-8 migrates as a 55/53 kDa (pro-form), 41/42 kDa (a cleaved/active or intermediate form), and 18 kDa (active form). The proform (55/53 kDa) is still seen in treated cells because not all cells undergo apoptosis at once.
2. May we ask if NB600-1343, NB100-56527 and NB500-208 can recognize both active form and profor
   • Product NB100-56527 recognises both pro and active forms of Caspase 8. NB500-208 recognises only the active form, and NB600-1343 has not been tested for specificity at this time.

Bioinformatics

• Gene Symbol: CASP8