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Caspase 8 (CASP8) ELISA Kit

INTENDED USE

Human caspase 8 ELISA kit is intended for determining in vitro quantitative amounts of caspase-8 (CASP8) in human tissue homogenates, plasma, serum and many biological fluids. This assay has a minimum sensitivity limit of 0.094 ng/ml.

BACKGROUND

Caspase 8 (CASP-8, CASP8) is a member of family of conserved cysteine-aspartic acid proteinases which have an important role in the regulation of apoptosis and cell death. This protein is able to induce Fas and a number of other apoptotic stimuli in order to carry out programmed cell death. The process of apoptosis is essentially induced via the tumor necrosis factor receptor 1 (TNFR1) and Fas/APO1 (CD95) receptor, this leads to the activation of caspase 8 which causes the release of the active fragments p10 and p18. This results in the activation of other downstream effector caspases (notably 1, 3, 6 and 7), it is caspase 3 which is responsible for eliciting the common morphological hallmarks of apoptosis such as cell shrinkage and DNA fragmentation.

There are evidence for many cancer therapies which are in development that involve using molecules that trigger the activation of Caspase-8 mediated apoptosis, especially in cancer cells. There are other therapies that use compounds which are able to disrupt cancer-specific mechanisms in order to inhibit caspase-8 mediated apoptosis.

Due to the fact that the activation of caspase-8 occurring at a such early stage of the apoptosis pathway, the detection of caspase-8 provides a useful alternative to measurement of apoptosis using Annexin V-FITC / PI, that measure phosphatidylserine (PS) present on the surface of apoptotic cells.

CASPASE 8 ELISA KIT CONTENT

All reagents supplied need to be stored at 2 °C – 8 °C, unopened reagents will retain reactivity until expiration date. Do not use reagents beyond this date.

  • One 96-Well Plate: Pre-coated with anti CASP8 antibody.
  • Standards: Concentrations 0, 0.156, 0.312, 0.625, 1.25, 2.5, 5.0, 10 ng/ml, made up from lyophilized recombinant human CASP8.
  • Sample/Standard Dilution Buffer.
  • Biotinylated-labelled CASP8 Antibody.
  • Antibody Dilution Buffer.
  • HRP-Streptavidin Conjugate (SABC).
  • SABC Dilution Buffer.
  • TMB Substrate.
  • Wash Buffer (25x).
  • Plate Sealer.
  • Product Instructions.

SENSITIVITY

Minimum detection limits of a caspase-8 (CASP8) using this human caspase 8 ELISA kit was 0.094 ng/ml. The dynamic range for this assay is 0.156 – 10.0 ng/ml.

ASSAY CHARACTERISTICS

– Caspase-8 (CASP8): ELISA
– Specificity: Highly specific for CASP8, no cross reactivity or interference between CASP8 and analogues was detected.
– Recovery: Serum (86 – 105%), EDTA Plasma (86 – 105%), Heparin Plasma (90 – 102%).
– Linearity: Serum (86 – 95%), EDTA Plasma (89 – 101%), Heparin Plasma (81 – 93%).
– Precison Intra-Assay: CV < 8%.
– Precison Inter-Assay: CV < 10%.
– Stability: Less than 10%.

REFERENCES

  1. Evolutionary analyses of caspase-8 and its paralogs: Deep origins of the apoptotic signaling pathways. Bioessays. (2015) 37 (7): 767-76. Review. Sakamaki K., et al.
  2. Old, new and emerging functions of caspases. Cell Death Differ. (2015) 22 (4): 526-39. Review. Shalini S., et al.
  3. Caspase-8 in apoptosis: the beginning of “the end”? IUBMB Life. (2000) 50 (2): 85-90. Review. Kruidering M. and Evan G.I.
  4. Caspases Connect Cell-Death Signaling to Organismal Homeostasis. Immunity. (2016) 44 (2): 221-31. Review. Galluzzi L., et al.
  5. Caspase-8 as a therapeutic target in cancer. Cancer Lett. (2013) 332 (2): 133-40. Review. Stupack D.G.

ADDITIONAL INFORMATION

  • Full Name: Caspase 8 (CASP8) ELISA Kit
  • Reactivity: Human
  • Sample Type: Serum, Tissue Homogenates, Plasma, Other Biological Fluids
  • Sensitivity: 0.094 ng/ml

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