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Cathepsin B ELISA Kit

INTENDED USE

Human cathepsin B ELISA kit is a protocol intended for detecting in vitro quantitative amounts of cathepsin B (CTSB) in human serum, plasma, biological fluids and tissue homogenates. This assay has a minimum detection limit of 0.094 ng/ml.

BACKGROUND

Cathepsin B belongs to family of lysosomal cysteine proteases and these are known to play vital functions during intracellular proteolysis. It can be synthesized on the rough endoplasmic reticulum as a pre-proenzyme that is 339 amino acid in length and it also possess a signal peptide that is 17 amino acids, this pro-cathepsin B of 43/46 kDa can then be transported to the Golgi apparatus where cathepsin B is formed. The mature cathepsin B consists of a heavy chain of 25-26 kDa and a light chain that is 5kDa which are linked by a dimer of disulphide.

Cathepsin B has the ability to enhance the activity of other protease such as urokinase, cathepsin D and matrix metalloproteinase, therefore giving it an important role in intercellular communication disruption, reducing protease inhibitor expression and in the proteolysis of extracellular matrix components. There are also evidence for its involvement in cannibalism and autophagy, this is advantageous in specific immune resistance and tumor malignancy.

Upregulation of cathepsin B has also be discovered in pre-malignant lesions and various other pathological conditions such as cancers. It is often proposed as a potentially effective biomarker for a number of cancers and its overexpression has been correlated with many metastatic and invasive cancers. There are a wide array of diseases which have displayed elevated levels of cathepsin B and this results in a number of different pathological processes such as production of toxic peptides, inflammation and cell death.

CATHEPSIN B 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 CTSB 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 cathepsin B.
  • Sample/Standard Dilution Buffer.
  • Biotinylated-labelled CTSB Antibody.
  • Antibody Dilution Buffer.
  • HRP-Streptavidin Conjugate (SABC).
  • SABC Dilution Buffer.
  • TMB Substrate.
  • Wash Buffer (25x).
  • Plate Sealer.
  • Product Instructions.

SENSITIVITY

The minimum detection sensitivity level of cathepsin-B (CTSB) using this human cathepsin B ELISA kit was 0.094 ng/ml. The dynamic assay range for this kit is 0.156 – 10.0 ng/ml.

ASSAY CHARACTERISTICS

– Cathepsin B (CTSB): ELISA
– Specificity: Highly specific for CTSB, no cross reactivity or interference between CTSB and analogues was detected.
– Recovery: Serum (88 – 105%), EDTA Plasma (91 – 99%), Heparin Plasma (90 – 95%).
– Linearity: Serum (89 – 105%), EDTA Plasma (84 – 95%), Heparin Plasma (81 – 96%).
– Precison Intra-Assay: CV < 8%.
– Precison Inter-Assay: CV < 10%.
– Stability: Less than 10%.

REFERENCES

  1. Specialized roles for cysteine cathepsins in health and disease. J Clin Invest. (2010) 120 (10): 3421-31. Review. Reiser J., et al.
  2. Molecular regulation of human cathepsin B: implication in pathologies. Biol Chem. (2003) 384 (6): 845-54. Review. Yan S. and Sloane B.F.
  3. Cell surface complex of cathepsin B/annexin II tetramer in malignant progression. Biochim Biophys Acta. (2000) 1477 (1-2): 215-30. Review. Mai J., et al.
  4. Inhibitors of cathepsin B. Curr Med Chem. (2006) 13 (19): 2309-27. Review. Frlan R. and Gobec S.
  5. Macrophage derived cystatin B/cathepsin B in HIV replication and neuropathogenesis. Curr HIV Res. (2014) 12 (2): 111-20. Review. Rivera L.E., et al.

ADDITIONAL INFORMATION

  • Full Name: Cathepsin B ELISA Kit
  • Reactivity: Human
  • Sample Type: Plasma, Serum, Biological Fluids, Tissue Homogenates
  • Sensitivity: 0.094 ng/ml

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