Neuronal Tissue Dissociation Enzyme
Contents
1. San Diego CA 92121 Toll Free 888 428 0558 U S amp Canada 858 457 1919 www genlantis com 2006 Genlantis and Gene Therapy Systems Inc2. Genlantis A division of Gene Therapy Systems Inc eroPapati Enyme lt A Neuronal Tissue Dissociation Enzyme Related Products Quantity NeuroPrep Medium 100 ml NM100100 NeuroPure E18 Primary Rat Hippocampal Cells NeuroPure E18 Primary Rat Cortical Cells NeuroPure P8 Primary Rat Cerebellar Cells NeuroPure Primary E18 Midbrain Cells ee NeuroPapain Enzyme is shipped at room temperature and should be stored at 4 C upon receipt It is shipping amp Storage stable for 6 months when stored properly INTRODUCTION The NeuroPapain Enzyme is an optimized formulation of the enzyme papain that is ideal for enzymatic dissociation of neuronal tissues such as the NeuroPure and NeuroStem Primary Rat Neurons NeuroPapain Enzyme provides rapid and safe enzymatic digestion of primary rodent neuronal tissues when used in conjuction with NeuroPrep Medium Cat NM100100 Following treatment with NeuroPapain in NeuroPrep Medium neuronal tissues are more easily dissociated via mechanical trituration and the number of viable dissociated cells can be increased by up to 100 However please note that for assays performed within 4 days of plating some digestion of surface proteins is inevitable MATERIALS AND METHODS Note The following protocol has been validated for use with the NeuroPure Primary Rat Neuronal Cells Please refer to the NeuroPure protocols for complete details 1 Add 5 mg of NeuroPapain Enzyme into 2 5
3. ml of NeuroPrep Medium Mix at 37 C for 15 minutes to completely dissolve the NeuroPapain Sterilize this solution with a 0 2 um filter prior to utilizing for tissue digestion Use within 3 hours for best results 2 Prior to enzymatic treatment allow the neuronal tissue to settle for 15 30 minutes at 4 C Alternatively place the tube containing the tissue in a 50 ml tube and spin down the cells at 1 100 rpm 200xg for 1 minute Transfer the medium from the cell vial to a separate sterile tube while being careful not to remove any loose tissue pieces Save the medium for trituration following NeuroPapain treatment 3 Immediately add 2 ml of sterile NeuroPapain solution to the tissue containing tube and allow the neuronal tissue to incubate for 30 minutes at 30 C Swirl every two minutes by hand 4 Following incubation spin down the cells at 1 100 rpm 200xg for 1 minute Remove the NeuroPapain solution again being careful not to disturb or remove the tissue 5 Add 1 ml of shipping medium back to the neuronal cells Save the other 1 ml of shipping medium for Step 4 in the NeuroPure protocol Mike Vengrow 12 5 06 10 24 AM Deleted NOTE you don t need to use the TM mark each time you mention the product name the convention is to use the TM indication only the first time you mention the product on each page 6 Proceed to Step 3 in the NeuroPure protocol v MV03142007 Genlantis Page 1 of 1 10190 Telesis Court
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