Cathepsin D ELISA Kit (Human) (OKBB00303)

Catalog No: OKBB00303 (Formerly GWB-J4JVV9)

Size:96 Tests

Human CTSD 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 D ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for Cathepsin D has been precoated onto 96-well plates. Standards(NS0, L21-L412) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for Cathepsin D 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 D 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 (ng/mL)	1.24	3.37	6.73	1.38	3.51	6.82
  St. Dev.	0.035	0.108	0.236	0.047	0.13	0.28
  %CV	2.82	3.20	3.50	3.40	3.70	4.10

• 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 D ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. Human Cathepsin D specific-specific polyclonal 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 D 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 D is a protein that in humans is encoded by the CTSD gene. This proteinase is a member of the peptidase C1 family, having a specificity similar to but narrower than that of pepsin A. It is mapped to 11p15.5. The cDNA encodes a 412-amino acid protein with 20 and 44 amino acids in a pre- and prosegment, respectively. Cathepsin D is one of the lysosomal proteinases. It is ubiquitously expressed and is involved in proteolytic degradation, cell invasion, and apoptosis. Mutations in this gene are involved in the pathogenesis of several diseases, including breast cancer and possibly Alzheimer disease and it has been considered as a breast cancer tumor marker.
• 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: <10 pg/ml
• Predicted Homology Based on Immunogen Sequence: No detectable cross-reactivity with any other cytokine
• Reference: 1. Faust PL, Kornfeld S, Chirgwin JM (Sep 1985). "Cloning and sequence analysis of cDNA for human cathepsin D". Proc
  Natl Acad Sci U S A 82 (15): 4910–4. doi:10.1073/pnas.82.15.4910. PMC 390467. PMID 3927292.
  2. Wolf M, Clark-Lewis I, Buri C, Langen H, Lis M, Mazzucchelli L (April 2003). "Cathepsin D specifically cleaves the
  chemokines macrophage inflammatory protein-1 alpha, macrophage inflammatory protein-1 beta, and SLC that are
  expressed in human breast cancer". Am. J. Pathol. 162 (4): 1183–90. PMC 1851240. PMID 12651610.
• Specificity: Natural and recombinant human Cathepsin D
• Immunogen: Expression system for standard: NS0; Immunogen sequence: L21-L412
• Description: For quantitative detection of human Cathepsin D in cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA).

Target Info

• Gene Symbol: CTSD
• Gene Full Name: cathepsin D
• Alias Symbols: CATD_HUMAN, Cathepsin D heavy chain, Ceroid lipofuscinosis neuronal 10, CLN10, CPSD, ctsd, Lysosomal aspartyl protease
• NCBI Gene Id: 1509
• Protein Name: Cathepsin D
• Description of Target: Cathepsin D is a protein that in humans is encoded by the CTSD gene. This proteinase is a member of the peptidase C1 family, having a specificity similar to but narrower than that of pepsin A. It is mapped to 11p15.5. The cDNA encodes a 412-amino acid protein with 20 and 44 amino acids in a pre- and prosegment, respectively. Cathepsin D is one of the lysosomal proteinases. It is ubiquitously expressed and is involved in proteolytic degradation, cell invasion, and apoptosis. Mutations in this gene are involved in the pathogenesis of several diseases, including breast cancer and possibly Alzheimer disease and it has been considered as a breast cancer tumor marker.
• Uniprot ID: P07339
• Protein Accession #: NP_001900.1
• Nucleotide Accession #: NM_001909.4