Cathepsin B ELISA Kit (Human) (OKBB00301)

Catalog No: OKBB00301

Size:96 Tests

Human CTSB ELISA Kit standard curve

Product Info

• Predicted Species Reactivity: Human
• Application: ELISA
• ELISA Kit Detection Method: Colorimetric, OD450 nm
• ELISA Kit Duration: ~ 3 Hours
• ELISA Kit Principle: Aviva's human Cathepsin B ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A polyclonal antibody from goat specific for Cathepsin B has been precoated onto 96-well plates. Standards(NS0, R18-I339) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for Cathepsin B is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The absorbance of yellow is proportional to the human Cathepsin B amount of sample captured in plate.
• ELISA Kit Range: 156 pg/ml - 10,000 pg/ml
• ELISA Kit Reproducibility:

  	Intra-Assay Precision			Inter-Assay Precision
  Sample	1	2	3	1	2	3
  Mean (pg/mL)	1350	3080	6170	1452	3209	6124
  St. Dev.	68.9	150.9	357.9	113.3	256.7	520.5
  %CV	5.10	4.89	5.80	7.80	7.99	8.49

• ELISA Kit Component:
  • Lyophilized recombinant human E-Cadherin standard: 10 ng/tube x 2
  • Anti-human E-Cadherin Antibody Well Plate: 96 Wells
  • Sample diluent buffer: 30 mL
  • Biotinylated anti-human E-Cadherin antibody: 130 uL, dilution 1:100
  • Antibody diluent buffer: 12 mL
  • Avidin-Biotin-Peroxidase Complex (ABC): 130 uL, dilution 1:100
  • ABC diluent buffer: 12 mL
  • TMB color developing agent: 10 mL
  • TMB stop solution: 10 mL
• Additional Information: Range: 156pg/ml-10,000pg/ml
  Principle: Aviva’s human Cathepsin B ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. Human Cathepsin B specific-specific monoclonal antibodies were precoated onto 96-well plates. The human specific detection polyclonal antibodies were biotinylated. The test samples and biotinylated detection antibodies were added to the wells subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the human Cathepsin B amount of sample captured in plate.
  Notes: 1. To inspect the validity of experiment operation and the appropriateness of sample dilution proportion, pilot
  experiment using standards and a small number of samples is recommended.
  2. The TMB Color Developing agent is colorless and transparent before using, contact us freely if it is not the case.
  3. Before using the Kit, spin tubes and bring down all components to the bottom of tubes.
  4. Duplicate well assay is recommended for both standard and sample testing.
  5. Don’t let 96-well plate dry, for dry plate will inactivate active components on plate.
  6. Don’t reuse tips and tubes to avoid cross contamination.
  7. To avoid to use the reagents from different batches together.
  8. In order to avoid marginal effect of plate incubation due to temperature difference ( reaction may be stronger in the
  marginal wells), it is suggested that the diluted ABC and TMB solution will be pre-warmed in 37C for 30 min before
  using.
  Background: Cathepsin B is an enzymatic protein belonging to the peptidase or protease families. In humans, it is coded by the CTSB gene.1, 2 And this gene is mapped to chromosome 8p22.3 The protein encoded by this gene is a lysosomal cysteine proteinase composed of a dimer of disulfide-linked heavy and light chains, both produced from a single protein precursor. It is a member of the peptidase C1 family. Cathepsin B was once suspected as a candidate protease participating in the conversion of β-amyloid precursor protein into the amyloid plaques found in Alzheimer's disease patients. However, this function is now known to be mediated by BACE1 protease. It is now thought that cathepsin B can degrade β-amyloid precursor protein into harmless fragments. Thus, it is conceivable cathepsin B may play a pivotal role in the natural defense against Alzheimer's disease.4 Overexpression of cathepsin B has been associated with esophageal adenocarcinoma and other tumors. At least five transcript variants encoding the same protein have been found for this gene. The standard product used in this kit is recombinant human Cathepsin B with the molecular mass of 37KDa.
• Reconstitution and Storage: Store at 4C for 6 months, at -20C for 12 months. Avoid multiple freeze-thaw cycles (Shipped with wet ice.)
• Sample Type: cell culture supernates, serum and plasma (heparin, EDTA)
• Sensitivity: <5 pg/ml
• Predicted Homology Based on Immunogen Sequence: No detectable cross-reactivity with any other cytokine
• Reference: 1. Chan SJ, San Segundo B, McCormick MB, Steiner DF (October 1986). "Nucleotide and predicted amino acid
  sequences of cloned human and mouse preprocathepsin B cDNAs". Proc. Natl. Acad. Sci. U.S.A. 83 (20): 7721–5.
  2. Cao L, Taggart RT, Berquin IM, Moin K, Fong D, Sloane BF (February 1994). "Human gastric adenocarcinoma
  cathepsin B: isolation and sequencing of full-length cDNAs and polymorphisms of the gene". Gene 139 (2): 163–9.
  3. Wang, X.; Chan, S. J.; Eddy, R. L.; Byers, M. G.; Fukushima, Y.; Henry, W. M.; Haley, L. L.; Steiner, D. F.; Shows, T.
  B. : Chromosome assignment of cathepsin B (CTSB) to 8p22 and cathepsin H (CTSH) to 15q24-q25. (Abstract)
  Cytogenet. Cell Genet. 46: 710-711, 1987.
  4. Mueller-Steiner S, Zhou Y, Arai H, Roberson ED, Sun B, Chen J, Wang X, Yu G, Esposito L, Mucke L, Gan L
  (September 2006). "Antiamyloidogenic and neuroprotective functions of cathepsin B: implications for Alzheimer's
  disease". Neuron 51 (6): 703–14
• Specificity: Natural and recombinant human Cathepsin B
• Immunogen: Expression system for standard: NS0; Immunogen sequence: R18-I339
• Description: For quantitative detection of human Cathepsin B in cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA).

Target Info

• Gene Symbol: CTSB
• Gene Full Name: cathepsin B
• Alias Symbols: aAmyloid precursor protein secretase, APP secretase, APPS, CATB_HUMAN, Cathepsin B heavy chain, Cathepsin B1, CPSB, CTSB, Cysteine protease
• NCBI Gene Id: 1508
• Protein Name: Cathepsin B
• Description of Target: Cathepsin B is an enzymatic protein belonging to the peptidase or protease families. In humans, it is coded by the CTSB gene.1, 2 And this gene is mapped to chromosome 8p22.3 The protein encoded by this gene is a lysosomal cysteine proteinase composed of a dimer of disulfide-linked heavy and light chains, both produced from a single protein precursor. It is a member of the peptidase C1 family. Cathepsin B was once suspected as a candidate protease participating in the conversion of β-amyloid precursor protein into the amyloid plaques found in Alzheimer's disease patients. However, this function is now known to be mediated by BACE1 protease. It is now thought that cathepsin B can degrade β-amyloid precursor protein into harmless fragments. Thus, it is conceivable cathepsin B may play a pivotal role in the natural defense against Alzheimer's disease.4 Overexpression of cathepsin B has been associated with esophageal adenocarcinoma and other tumors. At least five transcript variants encoding the same protein have been found for this gene. The standard product used in this kit is recombinant human Cathepsin B with the molecular mass of 37KDa.
• Uniprot ID: P07858
• Protein Accession #: NP_001899.1
• Nucleotide Accession #: NM_001908.4