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Product Spotlight: New Protease Activity Assays  |
 Innozyme Cathepsin B Activity Assay Kit, Fluorogenic
Cat. No.: CBA001 (View Product Insert)
Format: 100 Tests
Assay Range: 0.09 - 100 ng/ml
Assay Time: 30 minutes
Sample Type: Cell lysates, tissue extracts, and biological fluids. Purified enzymes for inhibitor screening.
Kit Contents: Cathepsin B enzyme, calibration standard, cathepsin B substrate, assay buffer, cathepsin B inhibitor, reduction reagent, cell lysis buffer, microtiter plate, plate sealer, and a directional insert.
Comments: A sensitive fluorogenic assay for the measurement of cathepsin B activity (Excitation: ~360-380 nm; Emission: ~430-460 nm). Cathepsin B is a cysteine proteinase which has been associated with increased invasivessness and development of the malignant cell phenotype. Cathepsin B has also been implicated in inflammatory airway disease. No cross reactivity with cathepsin L, H, or S.
| | | Calibration curve generated by various dilutions of the free AMC standard included in the kit. | The activity of native cathepsin B (Cat. No. 219364) with Z-Arg-Arg AMC substrate in MES buffer, pH 6.0, in the presence of EDTA and cysteine. After incubation at 37°C for 30 min. the free AMC was measured at excitation 360 nm and emission 460 nm. |
Sample | CB activity
(Fluorescence units/mg total protein/30 min) | Liver | 34.33 | Kidney | 2.14 | Placenta | 28.90 |
Cathepsin B activity in human tissue extracts as measured with Cathepsin B Activity Assay
| InnoZyme™ Cathepsin D Immunocapture Activity Assay Kit, Fluorogenic
Cat. No.: CBA002 (View Product Insert)
Format: 96 test
Sensitivity: 0.2 ng/ml
Assay Time: 1.5 hours
Sample Type: Cell lysates, tissue extracts, and biological fluids. Purified enzymes for inhibitor screening.
Kit Contents: Coated microtiter plate, substrate, cathepsin D standard, assay buffer, sample buffer, plate wash buffer, plate sealer, and a directional insert.
Comments: A highly selective fluorometric assay for the quantitative in vitro detection of human cathepsin D activity. This assay uses a monoclonal antibody to capture the cathepsin D. Activity is detected with an internally quenched fluorescent substrate (excitation max.:~328 nm; emission max: ~393 nm). | | Sensitivity: The minimum detectable activity, determined as the concentration of cathepsin D at two standard deviations above the mean fluorescence unit values of ten zero diluent is 02 ng/ml.
| Cell lysate | Cell line description | RFU/mg total protein/60 min | HT 1080 | Human fibrosarcoma | 422.0 ± 39.6 | A431 | Human epidermoid carcinoma | 2318.7 ± 228.8 | MCF-7 | Human breast adenocarcinoma | 1102.0 ± 108.8 | HeLa | Human cervical adenocarcinoma | 243.7 ± 18.9 | HL-60 | Human myoblastic acute promyelocytic leukemia derived from peripheral blood | 260.7 ± 34.5 | U251 | Human glioma | 402.5 ± 48.6 |
| Cathepsin D Activity: Representation of Cathepsin D activity in human cell lysated assayed with CBA002. Cell lysates were prepared with CytoBuster™ Protein Extraction Reagent, Cat. No. 71009. Fluorescence reading: excitation 320 nm, emission 405 nm. |
InnoZyme™ Gelatinase Activity Assay Kit, Fluorogenic
Cat. No.: CBA003 (View Product Insert)
Format: 100 Tests
Sensitivity: 10 ng/ml
Assay Time: 2 - 6 hours
Sample Type: Cell lysates, tissue extracts, and biological fluids. Purified enzymes for inhibitor screening.
Kit Contents: Gelatinase substrate, pro-MMP-2 positive control, inhibitor, APMA, assay buffer, microtiterplate, plate sealer, and a directional insert.
Comments: A sensitive fluorogenic assay (Excitation max: ~325 nm; Emission max.: ~393 nm) for the measurement of gelatinases (MMP-2 and MMP-9). Also useful for the screening of gelatinase inhibitors. MMP-2 and MMP-9 have been reported to induce angiogenesis during carcinogenesis. | 
This chart illustrates typical results upon dilution of the pro-MMP-2 positive control followed by incubation with the substrate in activation buffer at 37°C for 3 hours.
|  | Gelatinase activity observed in biological samples. Serum samples were diluted 5-fold and the HT1080 cell culture supernatant was diluted 3-fold. Samples were incubated at 37°C for 3 hours. |
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