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QIA76 Live/Dead Double Staining Kit

QIA76
  
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      Overview

      Replacement Information

      Key Spec Table

      Detection Methods
      Fluorescence
      Description
      OverviewThis kit uses a cell-permeable green fluorescent Cyto-dye (Ex. max.: 488 nm; Em. max.: 518 nm) to stain live cells, and propidium iodide (Ex. max.: 488 nm; Em. max.: 615 nm) to stain dead cells. Stained live and dead cells can be visualized by fluorescence microscopy using a band-pass filter which detects FITC and rhodamine. Viable cells will stain only with the Cyto-dye, fluorescing green, whereas the dead cells will stain with both Cyto-dye (green) and propidium iodide (red), resulting in a yellow fluorescence. Note: 1 T = 1 test.
      Optimal staining conditions may vary among different cell types and should be determined empirically by the investigator.
      Catalogue NumberQIA76
      Brand Family Calbiochem®
      Materials Required but Not Delivered Fluorescent microscope with a band-pass filter that detects FITC and rhodamine
      Microscope slides and cover slips
      References
      ReferencesLuther, E., and Kamenstsky, L.A. 1996. Cytometry 23, 272.
      Frey. T., et al. 1995. Cytometry 21, 265.
      Product Information
      Detection methodFluorescence
      Form100 Tests
      FormatFluorescence microscopy
      Kit containsStaining Buffer, Cyto-Dye, Propidium Iodide, and a user protocol.
      Quality LevelMQ100
      Applications
      Biological Information
      Assay time0.5 h
      Sample TypeCell suspensions or adherent cells
      Species Reactivity
      • A Broad Range Of Species
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      R PhraseR: 36/37/38-46

      Irritating to eyes, respiratory system and skin.
      May cause heritable genetic damage.
      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® Live/Dead Double Staining Kit can be used to distinguish between live and dead cells, which is essential for the study of growth control and cell death.
      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. Following initial use of the Staining Buffer, refrigerate (4°C) the remaining buffer.
      Protect from Moisture Protect from moisture
      Do not freeze Ok to freeze
      Packaging Information
      Transport Information
      Supplemental Information
      Kit containsStaining Buffer, Cyto-Dye, Propidium Iodide, and a user protocol.
      Specifications
      Global Trade Item Number
      Catalogue Number GTIN
      QIA76 0

      Documentation

      Live/Dead Double Staining Kit SDS

      Title

      Safety Data Sheet (SDS) 

      Live/Dead Double Staining Kit Certificates of Analysis

      TitleLot Number
      QIA76

      References

      Reference overview
      Luther, E., and Kamenstsky, L.A. 1996. Cytometry 23, 272.
      Frey. T., et al. 1995. Cytometry 21, 265.
      User Protocol

      Revision15-March-2017 JSW
      Form100 Tests
      FormatFluorescence microscopy
      Detection methodFluorescence
      Speciesa broad range of species
      StorageUpon arrival, store the entire contents of the kit at -20°C. Following initial use of the Staining Buffer, refrigerate (4°C) the remaining buffer.
      Intended useThe Calbiochem® Live/Dead Double Staining Kit can be used to distinguish between live and dead cells, which is essential for the study of growth control and cell death.
      Principles of the assayThe Calbiochem® Live/Dead Double Staining Kit provides ready-to-use staining reagents to conveniently discriminate between live and dead cells. The kit utilizes Cyto-dye, a cell-permeable green flourescent dye (Ex/Em = 488/518 nm), to stain live cells. Dead cells can be easily stained by propidium iodide (PI), a non-permeable red flourescent dye (Ex/Em= 488/615) that can only enter the cell when there is membrane damage that results in permeabilization. Stained live and dead cells can be visualized by flourescence microscopy using a band-pass filter that detects FITC and rhodamine. Cyto-Dye has also been used to discriminate between apoptotic and non-apoptotic cells.
      Materials provided• Solution A (Kit Component No. JA1897): 1 vial, 50 µl, 1 mM Cyto-Dye
      • Solution B (Kit Component No. JA1898): 1 vial, 50 µl, 1 mg/ml Propidium Iodide (PI)
      • Staining Buffer (Kit Component No. JA1899): 1 vial, 50 ml
      Materials Required but not provided Fluorescent microscope with a band-pass filter that detects FITC and rhodamine
      Microscope slides and cover slips
      Reagent preparation• Preparation of Staining Solution: Prior to use, prepare enough Staining Solution for the number of cell samples being tested. For example, to prepare 1 ml Staining Solution, mix 1 µl Solution A and 1 µl Solution B in 1 ml Staining Buffer. Use a ratio of 100 µl Staining Solution per 1 x 105 cells.
      Detailed protocol1. Collect cells by centrifugation at 500 x g for 5 min and discard the supernatant. For analyzing adherent cells, grow the cells directly on a coverslip.
      2. Resuspend the cell pellet with Staining Solution. Use 100 µl for every 1 x 105 cells.
      3. Incubate for 15 min at 37°C. For analyzing adherent cells, stain the cells directly on the coverslip.
      4. Transfer an aliquot of the cell suspension to a glass slide and cover the cells with a glass coverslip. For analyzing adherent cells, invert the coverslip on a glass slide to visualize the cells.
      5. Analyze the cells immediately under a fluorescence microscope using a band-pass filter that detects flourescein and rhodamine. Live cells stain only the cell-permeable Cyto-Dye, flourescing green. Dead cells stain with both the cell-permeable Cyto-Dye and the non-permeable propidium iodide, fluorescing red; the overlay of green and red appears yellow.

      Note: The optimal staining conditions may vary among different cell types, so it is recommended that the optimal concentration of Solution A and B be determined for individual cell types.
      Registered TrademarksCalbiochem® is a registered trademark of EMD Millipore, Corp., Inc.
      InteractivePathways™ is a trademark of EMD Millipore Corp.