Papain - lyophilized - 10mg

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    Papain - lyophilized - 10mg

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    DESCRIPTION: Papain APPLICATION: Use for Chondrex, Inc.’s Glycosaminoglycans Assay Kit (Cat # 6022)

    QUANTITY: 10 mg

    FORM: Lyophilized powder SOURCE: Papaya 

    USAGE: Papain digestion protocol for glycosaminoglycans, DNA, and denatured collagen

    1. Prepare 0.05 M phosphate buffered saline (PBS) pH 6.5 containing 5 mM cysteine and 5 mM EDTA, then add papain to a concentration of 125 µg/ml just before use.

    2. Suspend 10 mg of a tissue sample in 1 ml of the papain solution in a cryovial tube or rubber gasket sealed screw-cap tube.

    3. Incubate at 60˚C for several hours or overnight. NOTE 1: Papain-resistant polymers in scaffolds which constitute samples must be destroyed for accurate results. For example, agarose is degraded by incubation at 70˚C for 10 minutes, and chitosan can be physically crushed. NOTE 2: Do not exceed 70˚C during incubation for a DNA assay, as DNA may denature and fail to bind with the fluorescent dye.

    4. Centrifuge at 10,000 rpm for 5 minutes at room temperature.

    5. Transfer the supernatant to a 1.5 ml microcentrifuge tube. NOTE: Use this supernatant immediately to assay DNA with at least 1:50 dilution; sample storage will result in loss of up to 50% of the original DNA fluorescence (1).

    6. Store at -80˚C for subsequent analysis.

    STORAGE: -20°C 

    STABILITY: 1 year

    NOTES: N/A

    REFERENCES: N/A