QIA63
MMP-2 ELISA Kit
72 kDa Gelatinase ELISA Kit, Matrix Metalloproteinase 2 ELISA Kit, Gelatinase A ELISA Kit
For general questions please contact our Customer Service:
EMD Chemicals, Inc.
480 S. Democrat Road
08027 Gibbstown
United States
Phone: 800 854 3417
Fax: 800 776 0999
E-mail: emdinfo@emdchemicals.com
03 September 2010
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A non-isotopic colorimetric assay for the in vitro assay of human MMP-2 protein in tissue culture media, serum, or plasma.
| Product information | |||
|---|---|---|---|
| Format | 96-well plate | ||
| Form | 34 or 41 Tests | ||
| Detection method | Colorimetric | ||
| Species reactivity | human | ||
| Avoid freeze/thaw | Yes | ||
| Sensitivity | 0.1 ng/ml | ||
| Assay range | 1.56 - 50 ng/ml | ||
| Assay time | 5.5 h | ||
| Sample type | Tissue culture medium, serum, or plasma | ||
| Kit contains | Anti-MMP-2 Coated 96-Well Plate, MMP-2 Standard, Detector Antibody, 400X Conjugate, Conjugate Diluent, Substrate, Sample Diluent, Plate Wash Concentrate, Stop Solution, Plate Sealers, and a user protocol. | ||
| Declaration | Not available for sale in Japan. | ||
| Store and ship information | |||
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Hazardous Materials Attention: Due to the nature of the Hazardous Materials in this shipment, additional shipping charges may be applied to your order. Certain sizes may be exempt from the additional hazardous materials shipping charges. Please contact your local sales office for more information regarding these charges. |
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| Storage | -20°C | ||
| Ship |
Dry Ice Only
Multiple Toxicity Values, refer to MSDS |
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| Safety information | |||
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| S Phrase |
S: 23-26-36/37/39-45 Do not breathe fumes. In case of contact with eyes, rinse immediately with plenty of water and seek medical advice. Wear suitable protective clothing, gloves and eye/face protection. In case of accident or if you feel unwell, seek medical advice immediately (show the label where possible). |
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| R Phrase |
R: 23/24/25-35-36/37/38-40-43 Toxic by inhalation, in contact with skin and if swallowed. Causes severe burns. Irritating to eyes, respiratory system and skin. Limited evidence of a carcinogenic effect. May cause sensitization by skin contact. |
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| Data | |||
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 The assay has a 30% percent clinical sensitivity (ability to recognize affected individuals) at a 95% clinical specificity (ability to recognize unaffected individuals).  Astrocytoma and HS578T cells were plated out at various cell densities into 96 plate wells. Cells were incubated at 37°C for 24 h. Supernatant was measured in the MMP-2 ELISA. MMP-2 levels were detectable at cell concentrations of 4 X 10<sup>3</sup> Astrocytoma cells/well and 0.6 X 10<sup>4 HS578T cells/well and above.  The lower limit of detection (LLD), commonly used to define sensitivity, was measured by assaying four replicates of zero eight times using two different lots of plates and two different lots of detector antibody. The grand mean signal and pooled standard deviation of zero was calculated. The grand mean of each standard (run in replicates of four in the eight assays) was used for the standard curve, and the response, mean signal of zero plus two standard deviations, read in dose from the standard curve is the LLD; that is, the smallest dose that is not zero with 95% confidence.  The pooled coefficients of variation (according to the formula of Henry et. al., 1974) and between assay coefficients of variation are plotted against MMP-2 concentrations. The pooled data were collected from samples run eight times using two different lots of plates and two different lots of detector antibody in replicates of three on two separate occasions.  The study tested dilution-recovery of 28 positive samples. Dilutions were run in the MMP-2 ELISA and the "found" doses were plotted against the "expected" doses to determine linearity of dilution. The slope is very close to one and the intercept is not significantly different from zero. These studies are consistent with the absence of cross-reacting and matrix effects such as pH, salts, and endogenous binders that interfere with the reagents used in the assay. <b>Note:</b> Ca = Cancer; NHS = Normal Human Sera; TC= Tissue Culture.  Levels of MMP-2 detected by the ELISA after immunoaffinity extraction of MMP-2 positive samples with a MMP-2 antibody that is not used in the ELISA and a control antibody.  Levels of a highly purified 72 kDa MMP-2 that exists in a stable, but non-covalent 1:1complex with TIMP-2 detected after neutralization with a MMP-2 antibody that is not used in the ELISA.  Levels of MMP-2 detected by the MMP-2 ELISA after 2-aminophenylmercuric acetate (APMA) treatment. APMA promotes the autocatalytic cleavage of the N-terminal prosequence of the latent 72-kDa enzyme to yield the active form of the enzyme. HT1080 and astrocytoma tissue culture supernatants, recombinant proMMP-2 and normal human serum samples were either untreated (-APMA) or incubated for four hours at 37°C with 2 mM APMA (+APMA) or a volume of DMSO to control for the DMSO base of APMA (Vehicle Ctrl).  Levels of MMP-2 detected by the ELISA after immunoaffinity extraction of MMP-2 positive samples with a MMP-2 antibody that is not used in the ELISA and a control antibody. |
