Cystatin C ELISA Kit (Mouse) (OKBB00305)

Catalog No: OKBB00305

Size:96 Tests

Mouse CST3 ELISA Kit standard curve

Product Info

• Predicted Species Reactivity: Mouse
• Application: ELISA
• ELISA Kit Detection Method: Colorimetric, OD450 nm
• ELISA Kit Duration: ~ 3 Hours
• ELISA Kit Principle: Aviva's mouse Cystatin C ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from rat specific for Cystatin C has been precoated onto 96-well plates. Standards(NS0, M1-A140) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for Cystatin C 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 mouse Cystatin C amount of sample captured in plate.
• ELISA Kit Range: 312 pg/ml - 20,000 pg/ml
• ELISA Kit Reproducibility:

  	Intra-Assay Precision			Inter-Assay Precision
  Sample	1	2	3	1	2	3
  Mean (ng/mL)	3.05	6.26	13.16	3.12	6.37	13.66
  St. Dev.	0.131	0.194	0.474	0.212	0.376	0.861
  %CV	4.29	3.09	3.60	6.79	5.90	6.30

• 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: 0.312ng/ml-20ng/ml
  Principle: Aviva’s mouse Cystatin C ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. Mouse Cystatin C specific-specific polyclonal antibodies were precoated onto 96-well plates. The mouse 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 mouse Cystatin C 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: Cystatin C or cystatin 3 (formerly gamma trace, post-gamma-globulin or neuroendocrine basic polypeptide), a protein encoded by the CST3 gene, was originally described as a constituent of normal cerebrospinal fluid (CSF) and of urine from patients with renal failure.1 Cystatin 3 has a low molecular weight (approximately 13.3 kilodaltons), and it is removed from the bloodstream by glomerular filtration in the kidneys. In humans, all cells with a nucleus (cell core containing the DNA) produce cystatin C as a chain of 120 amino acids. It is found in virtually all tissues and bodily fluids. Cystatin C, which belongs to the type II cystatin gene family, is a potent inhibitor of lysosomal proteinases2 (enzymes from a special subunit of the cell that break down proteins) and probably one of the most important extracellular inhibitors of cysteine proteases3 (it prevents the breakdown of proteins outside the cell by a specific type of protein degrading enzymes). Moreover, cystatin C is involved in network reorganization in the epileptic dentate gyrus.4 The standard product used in this kit is recombinant human Cystatin C, consisting of 120 amino acids with the molecular mass of 15KDa.
• 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, plasma (heparin, EDTA) and urine
• Sensitivity: <10 pg/ml
• Predicted Homology Based on Immunogen Sequence: No detectable cross-reactivity with any other cytokine
• Reference: 1. Grubb, A.; Lofberg, H. : Human gamma-trace, a basic microprotein: amino acid sequence and presence in the
  adenohypophysis. Proc. Nat. Acad. Sci. 79: 3024-3027, 1982.
  2. Pirttila, T. J.; Manninen, A.; Jutila, L.; Nissinen, J.; Kalviainen, R.; Vapalahti, M.; Immonen, A.; Paljarvi, L.; Karkola,
  K.; Alafuzoff, I.; Mervaala, E.; Pitkanen, A. : Cystatin C expression is associated with granule cell dispersion in
  epilepsy. Ann. Neurol. 58: 211-223, 2005.
  3. Abrahamson, M. : Human cysteine proteinase inhibitors: isolation, physiological importance, inhibitory mechanism,
  gene structure and relation to hereditary cerebral hemorrhage. Scand. J. Clin. Lab. Invest. 48 (suppl. 191): 21-31,
  1988.
  4. Pirttila, T. J.; Manninen, A.; Jutila, L.; Nissinen, J.; Kalviainen, R.; Vapalahti, M.; Immonen, A.; Paljarvi, L.; Karkola,
  K.; Alafuzoff, I.; Mervaala, E.; Pitkanen, A. : Cystatin C expression is associated with granule cell dispersion in
  epilepsy. Ann. Neurol. 58: 211-223, 2005.
• Specificity: Natural and recombinant mouse Cystatin C
• Immunogen: Expression system for standard: NS0; Immunogen sequence: M1-A140
• Description: For quantitative detection of mouse Cystatin C in cell culture supernatants, serum, plasma( heparin, EDTA) and urine.

Target Info

• Gene Symbol: CST3
• Gene Full Name: cystatin C
• Alias Symbols: ARMD11, bA218C14.4 (cystatin C), Cst3, Cystatin-3, Cystatin-C, CYTC_HUMAN, Gamma-trace, Neuroendocrine basic polypeptide, Post-gamma-globulin
• NCBI Gene Id: 13010
• Protein Name: Cystatin-C
• Description of Target: Cystatin C or cystatin 3 (formerly gamma trace, post-gamma-globulin or neuroendocrine basic polypeptide), a protein encoded by the CST3 gene, was originally described as a constituent of normal cerebrospinal fluid (CSF) and of urine from patients with renal failure. Cystatin 3 has a low molecular weight (approximately 13.3 kilodaltons), and it is removed from the bloodstream by glomerular filtration in the kidneys. In mouses, all cells with a nucleus (cell core containing the DNA) produce cystatin C as a chain of 120 amino acids. It is found in virtually all tissues and bodily fluids. Cystatin C, which belongs to the type II cystatin gene family, is a potent inhibitor of lysosomal proteinases (enzymes from a special subunit of the cell that break down proteins) and probably one of the most important extracellular inhibitors of cysteine proteases (it prevents the breakdown of proteins outside the cell by a specific type of protein degrading enzymes). Moreover, cystatin C is involved in network reorganization in the epileptic dentate gyrus.
• Uniprot ID: P21460
• Protein Accession #: NP_034106.2
• Nucleotide Accession #: NM_009976.4