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QIA56 MMP-9 ELISA Kit

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QIA56
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Tableau de caractéristiques principal

Species ReactivityDetection Methods
HColorimetric

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QIA56-1EA
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      Description
      Overview

      This product has been discontinued.



      A non-isotopic, sandwich ELISA for the in vitro measurement of human MMP-9 in tissue culture supernatant, serum, or plasma.

      Catalogue NumberQIA56
      Brand Family Calbiochem®
      SynonymsGelatinase B ELISA Kit, 92 kDa Gelatinase ELISA Kit, Matrix Metalloproteinase 9 ELISA Kit
      Application Data
      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.


      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.

      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.

      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.

      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 β (TGFβ) 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.
      Materials Required but Not Delivered 2-20 µl, 20-200 µl and 200-1000 µl precision pipettors with disposable tips
      Wash bottle or multichannel dispenser for washing
      500 ml graduated cylinder
      Deionized or distilled H2O
      Spectrophotometer capable of measuring absorbance in 96-well plates using dual wavelength of 450/595 nm or 450/540nm
      References
      ReferencesMassova, 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.
      Product Information
      Detection methodColorimetric
      DeclarationNot available for sale in Japan.
      Form34 or 41 Tests
      Format96-well plate
      Kit contains96-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.
      Biological Information
      Assay range0.625 - 20 ng/ml
      Assay time3.5 h
      Sample TypeTissue culture media, serum, plasma
      Species Reactivity
      • Human
      Physicochemical Information
      Sensitivity0.1 ng/ml
      Safety Information
      R PhraseR: 23/24/25-35-36-40-43

      Toxic by inhalation, in contact with skin and if swallowed.
      Causes severe burns.
      Irritating to eyes.
      Limited evidence of a carcinogenic effect.
      May cause sensitization by skin contact.
      S PhraseS: 26-36/37/39-45

      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).
      Product Usage Statements
      Intended useThe Calbiochem® MMP-9 ELISA Kit is a non-isotopic immunoassay for the in vitro quantitation of human MMP-9 protein in tissue culture medium, serum, and plasma.
      Storage and Shipping Information
      Ship Code Dry Ice Only
      Toxicity Multiple Toxicity Values, refer to MSDS
      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.
      Storage -20°C
      Storage ConditionsUpon arrival, store the entire contents at -20°C. Avoid multiple freeze/thaw cycles. Do not expose reagents to excessive light.
      Avoid freeze/thaw Avoid freeze/thaw
      Do not freeze Ok to freeze
      Supplemental Information
      Kit contains96-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.
      Global Trade Item Number
      Référence GTIN
      QIA56-1EA 04055977209112