QIA117 Senescence Detection Kit

QIA117
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      Overview

      Replacement Information

      Key Specifications Table

      Detection Methods
      Colorimetric

      Pricing & Availability

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      QIA117-1KIT
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          Description
          OverviewUseful for the histochemical detection of senescent cells. The failure of cells to become senescent may lead to tumor progression. Higher levels of β-Galactosidase is considered as a good marker for senescent cells. The mechanism of this is believed to be due to the increased lysosomal content of senescent cells, resulting in an increase of the lysosomal enzyme β-galactosidase. Cells can be observed using microscope.
          Catalogue NumberQIA117
          Brand Family Calbiochem®
          Materials Required but Not Delivered PBS
          DMF
          References
          ReferencesKurz, D.J., et al. 2000. J. Cell Sci. 113, 3613.
          Dimri, G.P., et al. 1995. Proc. Natl. Acad. Sci. USA 92, 9363.
          Product Information
          Detection methodColorimetric
          Form250 Tests
          Format12-well plate
          Kit containsFixative Solution, X-Gal, Staining Solution, Staining Supplement, and a user protocol.
          Applications
          Application ReferencesFunasaka, T., et al. 2007. J. Biol. Chem. 282, 36362. Chen, Z., et al. 2005. Nature 436, 725.
          Biological Information
          Assay time30 min. plus overnight incubation
          Sample TypeIntact cells
          Physicochemical Information
          Dimensions
          Materials Information
          Toxicological Information
          Safety Information according to GHS
          Safety Information
          Product Usage Statements
          Intended useThe Calbiochem® Senescence Detection Kit is useful for the histochemical detection of senescent cells. The failure of cells to become senescent may lead to tumor progression. The mechanism of this is believed to be the increased lysosomal content of senescent cells resulting in an increase of the lysosomal enzyme β-galactosidase.
          Storage and Shipping Information
          Ship Code Dry Ice Only
          Toxicity Multiple Toxicity Values, refer to MSDS
          Storage -20°C
          Storage ConditionsUpon arrival store the entire contents of the kit at -20°C.
          Protect from Light Protect from light
          Do not freeze Ok to freeze
          Packaging Information
          Transport Information
          Supplemental Information
          Kit containsFixative Solution, X-Gal, Staining Solution, Staining Supplement, and a user protocol.
          Specifications

          Documentation

          Senescence Detection Kit SDS

          Title

          Safety Data Sheet (SDS) 

          Senescence Detection Kit Certificates of Analysis

          TitleLot Number
          QIA117

          References

          Reference overview
          Kurz, D.J., et al. 2000. J. Cell Sci. 113, 3613.
          Dimri, G.P., et al. 1995. Proc. Natl. Acad. Sci. USA 92, 9363.
          User Protocol

          Revision07-August-2008 JSW
          Form250 Tests
          Format12-well plate
          Detection methodColorimetric
          StorageUpon arrival store the entire contents of the kit at -20°C.
          Intended useThe Calbiochem® Senescence Detection Kit is useful for the histochemical detection of senescent cells. The failure of cells to become senescent may lead to tumor progression. The mechanism of this is believed to be the increased lysosomal content of senescent cells resulting in an increase of the lysosomal enzyme β-galactosidase.
          BackgroundSenescence is thought to be a tumor suppressive mechanism and an underlying cause of aging. It represents an arrested state in which the cells remain viable, but not stimulated to divide by serum or by passage in culture. Senescent cells display an increase in cell size, pH-dependent expression of β-galactosidase (β-Gal) activity, and altered patterns of gene expression.
          Principles of the assayThe Calbiochem® Senescence Detection Kit is designed to histochemically detect β-Gal activity in cultured cells and in tissue sections at pH 6.0, which is a known characteristic of senescent cells. β-Gal at pH 6.0 is present only in senescent cells and is not found in presenescent, quiescent, or immortal cells.
          Materials provided• Fixative Solution (Kit Component No. JA7633-125ML): 1 vial, 125 ml
          • X-Gal (Kit Component No. JA7634-150MG): 1 vial, 150 mg
          • Staining Solution (Kit Component No. JA7635-125ML): 1 vial, 125 ml
          • Staining Supplement (Kit Component No. JA7636-1.5ML): 1 vial, 1.5 ml
          Materials Required but not provided PBS
          DMF
          Reagent preparation• PBS: Prepare a 1X PBS solution (not provided). Use at least 3 ml per well. • X-gal: Dissolve 20 mg X-gal in 1 ml of DMF (N-N-dimethylformamide, not provided) to prepare a 20X stock solution. Aliquot and freeze (-20°) the unused X-gal solution. This stock solution is stable for up to 1 month at -20°C (protect from light). Always store X-gal in a polypropylene or glass container. Do not use polystyrene. • Staining Solution Mix: While cells are incubating in Fixative Solution (step 2 below), prepare Staining Solution Mix in a polypropylene plastic tube by adding 470 µl of Staining Solution, 5 µl Staining Supplement, and 25 µl of 20 mg/ml X-gal in DMF.
          Detailed protocolStaining

          Note: this procedure is described for one well of a 12-well culture plate. For a large plate, increase the volume proportionally. Please review the entire procedure before starting experiments.

          1.Remove culture medium and wash cells once with 1 ml 1X PBS.
          2. Fix cells with 0.5 ml of Fixative Solution at room temperature for 10 to 15 min.
          3. Wash the cells twice with 1 ml 1X PBS.
          4. Add 0.5 ml Staining Solution Mix to each well. Incubate at 37°C overnight.
          5. Observe cells under a microscope for development of blue color (approximately 200X total magnification).
          6. For long-term storage of the stained plates, remove the staining solution mix and overlay the cells with 70% glycerol. Store at 4°C.
          Application referencesFunasaka, T., et al. 2007. J. Biol. Chem. 282, 36362. Chen, Z., et al. 2005. Nature 436, 725.
          Registered TrademarksCalbiochem® is a registered trademark of EMD Chemicals, Inc.
          Interactive Pathways™ is a trademark of EMD Chemicals, Inc.