MMP-2 ELISA Kit (Mouse) (OKBB00221)

Catalog No: OKBB00221

Size:96 Tests

Mouse MMP2 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 MMP-2 ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from rat specific for MMP-2 has been precoated onto 96-well plates. Standards(NS0, A30-C662) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for MMP-2 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 MMP-2 amount of sample captured in plate.
• ELISA Kit Range: 625 pg/ml - 40,000 pg/ml
• ELISA Kit Reproducibility:

  	Intra-Assay Precision			Inter-Assay Precision
  Sample	1	2	3	1	2	3
  Mean (pg/mL)	779	3542	15328	844	4217	13668
  St. Dev.	41.28	170.01	950.33	54.01	231.93	997.76
  %CV	5.29	4.79	6.19	6.39	5.49	7.29

• ELISA Kit Component:
  • Lyophilized recombinant mouse MMP-2 standard: 10 ng/tube x 2
  • Anti-mouse MMP-2 Antibody Well Plate: 96 Wells
  • Sample diluent buffer: 30 mL
  • Biotinylated anti-mouse MMP-2 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 mouse MMP-2 ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. Mouse MMP-2 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 MMP-2 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: Type IV collagenase, 72-kD, is officially designated matrix metalloproteinase-2 (MMP2). It is also known as gelatinase, 72-kD. MMP-2 plays an essential role in angiogenesis and arteriogenesis, two processes critical to restoration of tissue perfusion after ischemia. MMP-2 expression is increased in tissue ischemia, but the responsible mechanisms remain unknown.¹ Matrix metalloproteinases (MMPs) catalyze extracellular matrix degradation. Control of their activity is a promising target for therapy of diseases characterized by abnormal connective tissue turnover. MMPs are expressed as latent proenzymes that are activated by proteolytic cleavage that triggers a conformational change in the propeptide (cysteine switch). The structure of proMMP-2 reveals how the propeptide shields the catalytic cleft and that the cysteine switch may operate through cleavage of loops essential for propeptide stability.^{2 The gene is localized to 16q21 using somatic cell hybrids and in situ hybridization.3 The standard product used in this kit is recombinant mouse MMP-2, consisting of 662 amino acids with the molecular mass of 72KDa. The detected MMP-2 includes zymogen and active enzyme.}
• 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: <10 pg/ml
• Predicted Homology Based on Immunogen Sequence: No detectable cross-reactivity with any other cytokine.
• Reference: 1. Lee, J. G.; Dahi, S.; Mahimkar, R.; Tulloch, N. L.; Alfonso-Jaume, M. A.; Lovett, D. H.; Sarkar, R. Intronic regulation
  of matrix metalloproteinase-2 revealed by in vivo transcriptional analysis in ischemia. Proc. Nat. Acad. Sci. 102:
  16345-16350, 2005.
  2. Morgunova, E.; Tuuttila, A.; Bergmann, U.; Isupov, M.; Lindqvist, Y.; Schneider, G.; Tryggvason, K. Structure of
  human pro-matrix metalloproteinase-2: activation mechanism revealed. Science 284: 1667-1670, 1999.
  3. Huhtala, P.; Eddy, R. L.; Fan, Y. S.; Byers, M. G.; Shows, T. B.; Tryggvason, K. Completion of the primary structure
  of the human type IV collagenase preproenzyme and assignment of the gene (CLG4) to the q21 region of
  chromosome 16. Genomics 6: 554-559, 1990.
• Specificity: Natural and recombinant mouse total MMP-2
• Immunogen: Expression system for standard: NS0; Immunogen sequence: A30-C662
• Description: For quantitative detection of mouse MMP-2 in cell culture supernatants, serum and plasma(heparin).

Target Info

• Gene Symbol: MMP2
• Gene Full Name: matrix metallopeptidase 2 (gelatinase A, 72kDa gelatinase, 72kDa type IV collagenase)
• Alias Symbols: 72 kDa gelatinase, 72 kDa type IV collagenase, CLG 4, CLG 4A, CLG4, CLG4A, Collagenase type IV A, EC 3.4.24.24, Gelatinase A, Gelatinase alpha, Gelatinase neutrophil, Matrix metallopeptidase 2 (gelatinase A, 72kDa gelatinase, 72kDa type IV collagenase), Matrix metalloproteinase 2, Matrix metalloproteinase 2 (gelatinase A, 72kD gelatinase, 72kD type IV collagenase), Matrix metalloproteinase II, Matrix metalloproteinase-2, MMP 2, MMP II, MMP-2, MMP2, MMP2_HUMAN, MONA, Neutrophil gelatinase, PEX, TBE 1, TBE-1
• NCBI Gene Id: 17390
• Protein Name: 72 kDa type IV collagenase
• Description of Target: Type IV collagenase, 72-kD, is officially designated matrix metalloproteinase-2 (MMP2). It is also known as gelatinase, 72-kD. MMP-2 plays an essential role in angiogenesis and arteriogenesis, two processes critical to restoration of tissue perfusion after ischemia. MMP-2 expression is increased in tissue ischemia, but the responsible mechanisms remain unknown.1 Matrix metalloproteinases (MMPs) catalyze extracellular matrix degradation. Control of their activity is a promising target for therapy of diseases characterized by abnormal connective tissue turnover. MMPs are expressed as latent proenzymes that are activated by proteolytic cleavage that triggers a conformational change in the propeptide (cysteine switch). The structure of proMMP-2 reveals how the propeptide shields the catalytic cleft and that the cysteine switch may operate through cleavage of loops essential for propeptide stability.2 The gene is localized to 16q21 using somatic cell hybrids and in situ hybridization.3 The standard product used in this kit is recombinant mouse MMP-2, consisting of 662 amino acids with the molecular mass of 72KDa. The detected MMP-2 includes zymogen and active enzyme.
• Uniprot ID: P33434
• Protein Accession #: NP_032636.1
• Nucleotide Accession #: NM_008610.3