MMP-13 ELISA Kit (Human) (OKBB00228)

Catalog No: OKBB00228 (Formerly GWB-ZZD182 )

Size:96 Tests

Human MMP13 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 MMP13 ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for MMP13 has been precoated onto 96-well plates. Standards(NS0, L20-C471) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for MMP13 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 MMP13 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)	199	723	3701	258	830	4198
  St. Dev.	10.547	37.59	218.35	15.738	54.78	247.682
  %CV	5.3	5.19	5.89	6.1	6.6	5.9

• ELISA Kit Component:
  • Lyophilized recombinant human MMP-7 standard: 10 ng/tube x 2
  • Anti-human MMP-7 Antibody Well Plate: 96 Wells
  • Sample diluent buffer: 30 mL
  • Biotinylated anti-human MMP-7 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: 78pg/ml => 5000pg/ml
  Principle: Aviva’s human MMP13 ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. Human MMP13 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 MMP13 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: Collagenase 3 is an enzyme that in humans is encoded by the MMP13 gene.[1][2]Most MMP's are secreted as inactive proproteins which are activated when cleaved by extracellular proteinases. The protein encoded by this gene cleaves type II collagen more efficiently than types I and III. [3] Mitchell et al. (1996) concluded that the expression of MMP13 in osteoarthritic cartilage and its activity against type II collagen indicates that the enzyme plays a significant role in cartilage collagen degradation and must, therefore, form part of a complex target for proposed therapeutic interventions based on collagenase inhibition. Reboul et al. (1996) likewise presented data on collagenase-3 expression and synthesis in human cartilage cells and suggested its involvement in human osteoarthritis cartilage pathophysiology. It may be involved in articular cartilage turnover and cartilage pathophysiology associated with osteoarthritis. The gene is part of a cluster of MMP genes which localize to chromosome 11q22.3.
• 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)
• Sensitivity: <5 pg/ml
• Predicted Homology Based on Immunogen Sequence: No detectable cross-reactivity with any other cytokine.
• Reference: 1. Freije JM, Diez-Itza I, Balbin M, Sanchez LM, Blasco R, Tolivia J, Lopez-Otin C (Jul 1994). "Molecular cloning
  and expression of collagenase-3, a novel human matrix metalloproteinase produced by breast carcinomas". J
  Biol Chem 269 (24): 16766–73.
  2. "Entrez Gene: MMP13 matrix metallopeptidase 13 (collagenase 3)".
  3. "Entrez Gene: MMP13 matrix metallopeptidase 13 (collagenase 3)".
• Specificity: Natural and recombinant human total MMP13
• Immunogen: Expression system for standard: NS0; Immunogen sequence: L20-C471
• Description: For quantitative detection of human MMP13 in cell culture supernatants, serum and plasma( heparin).

Target Info

• Gene Symbol: MMP13
• Gene Full Name: matrix metallopeptidase 13 (collagenase 3)
• Alias Symbols: CLG 3, CLG3, Collagenase 3, Collagenase3, MANDP1, Matrix metalloproteinase 13, Matrix metalloproteinase-13, MMP 13, MMP-13, Mmp13, MMP13_HUMAN, UMRCASE
• NCBI Gene Id: 4322
• Protein Name: Collagenase 3
• Description of Target: Collagenase 3 is an enzyme that in humans is encoded by the MMP13 gene.[1][2]Most MMP's are secreted as inactive proproteins which are activated when cleaved by extracellular proteinases. The protein encoded by this gene cleaves type II collagen more efficiently than types I and III. [3] The expression of MMP13 in osteoarthritic cartilage and its activity against type II collagen indicates that the enzyme plays a significant role in cartilage collagen degradation and must, therefore, form part of a complex target for proposed therapeutic interventions based on collagenase inhibition. It may be involved in articular cartilage turnover and cartilage pathophysiology associated with osteoarthritis. The gene is part of a cluster of MMP genes which localize to chromosome 11q22.3.
• Uniprot ID: P45452
• Protein Accession #: NP_002418.1
• Nucleotide Accession #: NM_002427.3