QIA117 Senescence Detection Kit

QIA117
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Overview

Replacement Information

Key Specifications Table

Detection Methods
Colorimetric

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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.
      Quality LevelMQ100
      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.