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MD Bioproducts/NeuroFreeze/M036041/10 ml or 30 mL

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货号:M036041
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品牌:MD Bioproducts
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商品描述
Overview: 

TheNeuroFreezekitcanbeusedtofreezeprimaryneuronsfromE18/19ratandP0/P1mousehippocampiandcortices.NeuronsfrozeninthismediamaintainhighlevelsofviABIlity(upto90%)uponthawing.Theseneuronsshowalltheexpectedcharacteristicsofprimaryneurons,includingappropriatemorphologyandexpressionofneuronalandsynapticMarkers.Theavailabilityofthismediawillenableinvestigatorstofreezeprimaryneuronsfromembryonicrats(E18)orP0/P1wildtypeorgeneticallyengineeredmice.

Data/Specifications: 

Species: mouse,rat

 

 

 

How_To_Use:

FreezingofPrimaryNeurons:    

  1. Obtaindissociatedprimaryneuronsfromrat/mousehippocampus/cortexasdescribed(Beaudoinetal).Re-sUSPenddissociatedcellsinminimalvolumeofplatingmedia.
  2. EstimatecellviabilityonaHemocytometer.
  3. Donotexceedamaximumof4millioncells/vialforfreezing.Itisrecommendedtofreeze1to4millioncellspervial.
  4. Re-suspendcellstobefrozenin900μlofReagentA.
  5. Add10%(100μl for1mL)ofReagentBtothevial.
  6. Add1%(10μlfor1mL)ofReagentCtothevial.
  7. Gentlymixandplacethevialinisopropanol bathin-80°Cfreezer.
  8. Nextday,placevial(s)in liquidnitrogen.

THAWINGPROTOCOL

  1. Followyourlaboratory’sprotocolforcoatingsubstrate.
  2. Addpre‐maintenancemedia(ReagentD)todishes/platesandincubatein37°Cincubatorforaminimumof2hours(recommended4hours).
  3. Bringavialofcellsfromliquidnitrogenandquicklythawin37°Cwaterbath(usuallytakesabout90seconds)–itiscriticalthatthawingisaccomplishedasfastaspossIBLe.
  4. Estimatecellviability.
  5. Seedcellsontothesubstrate(SeePlatingDensityChartbelow)
  6. Aftercellshaveattachedtothesubstrate(3-4hours),replacepre-maintenancemediawithyourlab’sneuronmaintenancemedia(B-27containingmaintenancemediaisrecommended).
  7. MaintainneuronsinB27containingmaintenancemediaat37°C.

 

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