Caspase 8 antibody (Caspase 8, Apoptosis-Related Cysteine Peptidase) (AA 430-482)

Details for Product anti-CASP8 Antibody No. ABIN724205, Supplier: Log in to see
Antigen
  • casp8-A
  • xCaspase-8
  • casp8
  • CASP8
  • CASP-8
  • FLICE
  • MACH
  • Mch5
  • ALPS2B
  • CAP4
  • Casp-8
  • MCH5
  • caspase-8
  • zgc:92075
  • caspase 8, apoptosis-related cysteine peptidase
  • death related ced-3/Nedd2-like protein
  • caspase-8
  • caspase 8
  • casp8
  • Dredd
  • CASP8
  • AaeL_AAEL014148
  • LOC100222284
  • Casp8
  • LOC100348477
Alternatives
anti-Rat (Rattus) Caspase 8 antibody for ELISA
Epitope
AA 430-482
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Reactivity
Human, Mouse (Murine), Rat (Rattus)
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Host
Rabbit
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Clonality
Polyclonal
Conjugate
This Caspase 8 antibody is un-conjugated
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Application
Immunofluorescence (Cultured Cells) (IF (cc)), Immunofluorescence (Paraffin-embedded Sections) (IF (p)), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Western Blotting (WB)
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Options
Supplier
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Supplier Product No.
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Available images

'Independent Validation' Badge
Lot Number 131127
Method validated Western Blotting
Positive Control HeLa cells
Negative Control c6/36 mosquito cells (non-reactive species)
Notes A strong band was observed at the correct molecular weight in the positive control sample. No major bands were observed in the negative sample.
Primary Antibody
  • Antigen: Caspase 8, Apoptosis-Related Cysteine Peptidase (CASP8) (1:200 dilution)
  • Catalog number: ABIN724205
  • Supplier: Log in to see
  • Supplier catalog number: Log in to see
  • Lot number: 131127
Secondary Antibody
  • Antigen: Goat Anti-Rabbit IgG (H + L)-HRP Conjugate (1:20,000 dilution)
  • Supplier: Log in to see
  • Catalog number: Log in to see
  • Lot number: L170-6515
Controls
  • Positive control: HeLa cell extract
  • Negative control: c6/36 cell extract
Protocol
  • 1. Total protein extracts were boiled in 1X SDS Sample Buffer containing 1% SDS and 1.25% β-mercaptoethanol at 95°C for 5 min prior to loading.
  • 2. 20 μg of boiled extracts were loaded and resolved on 8-16% SDS-polyacrylamide gel.
  • 3. The Thermo Scientific - Spectra Multicolor Broad Range (Cat # 26634) were used as molecular mass markers.
  • 4. Proteins were then transferred onto PVDF membrane by wet transfer and protein transfer was confirmed with Ponceau-S staining.
  • 5. The PVDF membrane was incubated with 25 mL of blocking buffer [Tris Buffered Saline, pH 7.4 plus 0.1% TW20 (TBST)] containing 5% (W/V) BSA at room temperature for 1 h.
  • 6. The membrane was rinsed with TBST once.
  • 7. The membrane was immersed with the protein side up in the primary antibody solution (CASP8; 1:200) in TBST containing 5% (W/V) BSA and incubated for 16 h at 4°C.
  • 8. The membrane was rinsed in TBST thrice for 5 min each.
  • 9. The membrane was incubated in the HRP-conjugated secondary antibody solution (Goat anti-rabbit IgG-HRP; 1:20,000) in TBST containing 5% (W/V) BSA and incubated for 1 hour at room temperature (~26°C) with gentle agitation.
  • 10. The membrane was rinsed thrice TBST thrice for 5 min each.
  • 11. The membrane was rinsed in TBS twice for 30 s each.
  • 12. Signals were detected with ECL-2 Substrate. The blot was scanned for 300 s.
  • 13. The membrane was rinsed three times TBST.
  • 14. Incubated in Acidic Glycine Stripping Buffer at room temperature with gentle agitation for 3 times, 10 min each.
  • 15. The membrane was washed in TBST 2 times for 10 min each.
  • 16. Repeated Steps 5-12 with the loading control antibody (anti-Actin; 1:6,000) and its matching secondary antibody (Goat anti-rabbit IgG-HRP; 1:20,000).
Experimental Notes - Nothing to note.
Validation Images
Western Blotting validation image for anti-Caspase 8, Apoptosis-Related Cysteine Peptidase (CASP8) (AA 430-482) antibody (ABIN724205) Figure 1: Western blot of lysates from HeLa cells (Lane 1), and c6/36 cells (Lane 2) ...
Immunogen KLH conjugated synthetic peptide derived from mouse Caspase-8 subunit p10
Isotype IgG
Purification Purified by Protein A.
Alternative Name Caspase 8 (CASP8 Antibody Abstract)
Background

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 CASP1. 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.

Subcellular location: Cytoplasm

Synonyms: MACH, Mch5, FLICE, CASP-8, Caspase-8, Casp8

Gene ID 12370
UniProt O89110
Research Area Proteolysis / Ubiquitin, Apoptosis/Necrosis
Pathways Apoptosis, Caspase Cascade in Apoptosis, TLR Signaling, Activation of Innate immune Response, Tube Formation, Positive Regulation of Endopeptidase Activity, Toll-Like Receptors Cascades
Application Notes WB 1:100-1000
IHC-P 1:100-500
IF(IHC-P) 1:50-200
IF(ICC) 1:50-200
Restrictions For Research Use only
Format Liquid
Concentration 1 μg/μL
Buffer Aqueous buffered solution containing 1 % BSA, 50 % glycerol and 0.09 % sodium azide.
Preservative Sodium azide
Precaution of Use This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE, which should be handled by trained staff only.
Storage -20 °C
Storage Comment Store at -20°C
Expiry Date 12 months
Supplier Images
Immunohistochemistry (IHC) image for anti-Caspase 8 antibody (Caspase 8, Apoptosis-Related Cysteine Peptidase) (AA 430-482) (ABIN724205) Formalin-fixed and paraffin embedded human rectal carcinoma labeled with Anti-Caspase...
SDS-PAGE (SDS) image for anti-Caspase 8 antibody (Caspase 8, Apoptosis-Related Cysteine Peptidase) (AA 430-482) (ABIN724205) L1 mouse liver lysates L2 mouse spleen lysates probed with Anti Caspase 8 Polyclonal ...
Western Blotting (WB) image for anti-Caspase 8 antibody (Caspase 8, Apoptosis-Related Cysteine Peptidase) (AA 430-482) (ABIN724205) Image provided by the Independent Validation Program (badge number 29759). Lane 1: He...
Immunofluorescence (Cultured Cells) (IF (cc)) image for anti-Caspase 8 antibody (Caspase 8, Apoptosis-Related Cysteine Peptidase) (AA 430-482) (ABIN724205) Image provided by One World Lab validation program. MCF-7 cells probed with Rabbit An...
Immunofluorescence (Cultured Cells) (IF (cc)) image for anti-Caspase 8 antibody (Caspase 8, Apoptosis-Related Cysteine Peptidase) (AA 430-482) (ABIN724205) Image provided by One World Lab validation program. A431 cells probed with Rabbit Ant...
Product cited in: Tulsulkar, Glueck, Hinds, Shah: "Ginkgo biloba Extract Prevents Female Mice from Ischemic Brain Damage and the Mechanism Is Independent of the HO1/Wnt Pathway." in: Translational stroke research, Vol. 7, Issue 2, pp. 120-31, 2016 (PubMed).

Akinrinde, Oyagbemi, Omobowale, Asenuga, Ajibade: "Alterations in blood pressure, antioxidant status and caspase 8 expression in cobalt chloride-induced cardio-renal dysfunction are reversed by Ocimum gratissimum and gallic acid in Wistar rats." in: Journal of trace elements in medicine and biology : organ of the Society for Minerals and Trace Elements (GMS), Vol. 36, pp. 27-37, 2016 (PubMed). (Sample species: Rat (Rattus)).

Gao, Liu, Li, Wu, Peng, Jing: "Hispidulin induces mitochondrial apoptosis in acute myeloid leukemia cells by targeting extracellular matrix metalloproteinase inducer." in: American journal of translational research, Vol. 8, Issue 2, pp. 1115-32, 2016 (PubMed).

Daverey, Agrawal: "Curcumin alleviates oxidative stress and mitochondrial dysfunction in astrocytes." in: Neuroscience, Vol. 333, pp. 92-103, 2016 (PubMed).

Wang, Zhao, Yu, Feng, Zhang, Kou, Chu, Cui, Li, Zhang, Shen, Min: "Regulation of steroid hormones and energy status with cysteamine and its effect on spermatogenesis." in: Toxicology and applied pharmacology, Vol. 313, pp. 149-158, 2016 (PubMed).

Zhao, Zhang, Liu, Zhang, Hao, Li, Chen, Shen, Tang, Min, Meng, Wang, Yi, Zhang: "Hydrogen Sulfide and/or Ammonia Reduces Spermatozoa Motility through AMPK/AKT Related Pathways." in: Scientific reports, Vol. 6, pp. 37884, 2016 (PubMed). Method employed by authors: Western Blotting (WB) (Sample species: Pig (Porcine)).

Fang, Zhang, Qi, Fan, Luo, Liu, Tan: "Evodiamine induces G2/M arrest and apoptosis via mitochondrial and endoplasmic reticulum pathways in H446 and H1688 human small-cell lung cancer cells." in: PLoS ONE, Vol. 9, Issue 12, pp. e115204, 2014 (PubMed).

Wang, Han, Zhou, Wu, Wang, Jia, Lv, Cheng, Pan, Liu, Zhou, Sun: "Inhibition of hydrogen sulfide synthesis provides protection for severe acute pancreatitis rats via apoptosis pathway." in: Apoptosis : an international journal on programmed cell death, Vol. 18, Issue 1, pp. 28-42, 2013 (PubMed).

Wu, Tang, Jiang, Li, Jiang, Liu: "PCSK9 siRNA inhibits HUVEC apoptosis induced by ox-LDL via Bcl/Bax-caspase9-caspase3 pathway." in: Molecular and cellular biochemistry, Vol. 359, Issue 1-2, pp. 347-58, 2011 (PubMed).

Jeng, Dyson: "Direct measurement of the aspartic acid 26 pKa for reduced Escherichia coli thioredoxin by 13C NMR." in: Biochemistry, Vol. 35, Issue 1, pp. 1-6, 1996 (PubMed).