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 | -20°C |  | Dry ice |  | Hazardous |  | Multiple Toxicity Values, refer to MSDS | | Note: Store and Ship conditions may differ. | | See Key |
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92 kDa Gelatinase ELISA Kit
Gelatinase B ELISA Kit
Matrix Metalloproteinase 9 ELISA Kit | Format: 96-well plate | | Form: 34 or 41 Tests | | Detection Method: Colorimetric | | Species Reactivity: human | | Specificity: Human pro-MMP-9 protein and MMP-9/TIMP-1 complex | | Sensitivity: 0.1 ng/ml | | Assay Range: 0.625 - 20 ng/ml | | Assay Time: 3.5 h | | Sample Type: Tissue culture media, serum, plasma | | Kit Contents: 96-Well Coated Plate, MMP-9 Standard, Biotinylated Detector Antibody, 400X Streptavidin Peroxidase Conjugate, Diluent, Substrate, Assay Buffers, Stop Solution, Plate Sealer, and a user protocol. | | Comments: A non-isotopic, sandwich ELISA for the in vitro measurement of human MMP-9 in tissue culture supernatant, serum, or plasma. | | Ref.: Massova, I., et al. 1998. FASEB J. 12, 1075. Xie, B.,et al. 1998. J. Biol. Chem. 273, 11576. Borden, P., and Heller, R. A. 1997. Crit. Rev. Eukaryot. Gen. Expression 7, 159. Chambers, A. F., and Matrisian, L. M. 1997. J. Natl. Cancer Inst. 89, 1260. DeClerck, Y.A., et al. 1997. Adv. Exp. Med. Biol. 425, 89. Gomis-Ruth, F. X., et al. 1997. Nature 389, 77. Murphy, G., and Knauper, V. 1997. Matrix Biol. 15, 511. Olson, M. W., et al. 1997. J. Biol. Chem. 272, 29975. Woodhouse, F. C., et al. 1997. Cancer 80, 1529. Yu, A. E., et al. 1997. Drugs Aging 11, 229. Coussens, L. M., and Werb, Z. 1996. Chem. Biol. 3, 895. Lim Y.T., et al. 1996. J. Cell. Physiol. 167, 333. Kohn, E. C., and Liotta, L. A. 1995. Cancer Res. 55, 1856. Shapiro, S.D., et al. 1995. J. Biol. Chem. 270, 6351. Murphy, G., et al. 1994. J. Biol. Chem. 269, 6632. Fridman, R., et al. 1993. Biochem J. 289, 411. Goldberg, G. L., et al. 1992. J. Biol. Chem. 267, 4583. Morodomi T.,et al. 1992. Biochem. J. 285, 603. | | R: 23/24/25-35-36-40-43; S: 26-36/37/39-45 | | Not available for sale in Japan. |
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* Additional Product Information
Example Standard Curve. The mean signal of each standard run in replicates of four in eight assays using two different lots of plates and two different lots of detector antibody.
Levels of MMP-9 Following APMA Treatment. APMA promotes the autocatalytic cleavage of the N-terminal propeptide of the latent 92-kDa enzyme to yield the active form of the enzyme. Analysis of the samples by zymography showed both cleavage of the proenzyme by APMA and a good correlation with the levels of the proenzyme determined with the MMP-9 ELISA. Several methods of MMP-9 induction were used to generate positive samples such as treatment with 50 ng/ml amphiregulin (AR), 50 ng/ml epidermal growth factor (EGF), 1 ng/ml transforming growth factor b (TGFb) and 25 ng/ml phorbol 12-myristate 13-acetate (PMA). Normal human sera (NHS) containing moderate levels of MMP-9 were treated with APMA and analyzed.
Parallelism. The study tested dilution-recovery of ten positive samples. The dilutions were run in the MMP-9 ELISA Kit and the "found" doses were plotted against the "expected" doses to determine linearity of dilution. The slope is not significantly different from 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. Note: Ca = Cancer; NHS = Normal Human Sera; NHP = Normal Human Plasma; PMA = Phorbol 12-myristate 13-acetate; EGF = Epidermal Growth Factor; AR = Amphiregulin; Both = EGF and AR; A = 2 x 106 cells/ml; B = 1.0 x 106 cells/ml.
Precision. The pooled coefficients of variation (according to the formula of Henry et. al., 1974) and between assay coefficients of variation are plotted against MMP-9 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 four on two separate occasions.
Sensitivity.
Specificity. Levels of MMP-9 detected by the ELISA after immunoaffinity extraction of MMP-9 positive samples (PMA treated HT1080 and HL-60 cells) with a MMP-9 antibody that is not used in the ELISA, a TIMP-1 antibody, a TIMP-2 antibody and a TIMP-3 antibody.
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