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1、LABORATORYSTOCKSOLUTIONSANDAPPENDIX2EQUIPMENTCommonStockSolutions,Buffers,andMediaAPPENDIX2ARECIPESThissectiondescribesthepreparationofbuffersandreagentsusedinthismanualforcellculture,manipulationoftissue,andcellbiologicalmethods.Whenpreparingsolutions,usedeionizedo
2、rdistilledwaterandreagentsofthehighestavailablegrade.Sterilization—byfiltrationthrougha0.22-mmfilterorbyautoclaving—isrecommendedformostsolutionsstoredatroomtemperatureandisessentialforcellcultureapplications.Wherestorageconditionsarenotspecified,storeupto6monthsatr
3、oomtemperature.Discardanyreagentthatshowsevidenceofcontamination,precipitation,ordiscoloration.Acid,concentratedstocksolutionsSeeTableA.2A.1.Acidprecipitationsolution1MHCl0.1MsodiumpyrophosphateNucleicacidscanalsobeprecipitatedwitha10%(w/v)solutionoftrichloroacetica
4、cid(TCA);however,thisrecipeischeaper,easiertoprepare,andjustasefficacious.Ammoniumhydroxide,concentratedstocksolutionSeeTableA.2A.1.Ammoniumacetate,10MDissolve385.4gammoniumacetatein150mlH2OAddH2Oto500mlAmmoniumsulfate,saturated76gammoniumsulfate100mlH2OHeatwithstir
5、ringtojustbelowboilingpointLetstandovernightatroomtemperatureTableA.2A.1MolaritiesandSpecificGravitiesofConcentratedAcidsandBasesa1MMolecularMolaritySpecificAcid/base%byweightsolutionweight(approx.)gravity(ml/liter)Aceticacid(glacial)60.0599.617.41.0557.5Ammoniumhyd
6、roxide35.02814.80.9067.6Formicacid46.039023.61.20542.49825.91.2238.5Hydrochloricacid36.463611.61.1885.9Nitricacid63.017015.71.4263.7Perchloricacid100.46609.21.54108.87212.21.7082.1Phosphoricacid98.008514.71.7067.8Sulfuricacid98.079818.31.83554.5aLaboratoryStockCAUTI
7、ON:Handlestrongacidsandbasescarefully.SolutionsandEquipmentCurrentProtocolsinCellBiology(1998)A.2A.1-A.2A.10A.2A.1Copyright©1998byJohnWiley&Sons,Inc.ATP,100mM1gATP(adenosinetriphosphate)12mlH2OAdjustpHto7.0with4MNaOHAdjustvolumeto16.7mlwithH2OStoreinaliquotsindefini
8、telyat-20°CBase,concentratedstocksolutionsSeeTableA.2A.1.BSA(bovineserumalbumin),10%(w/v)Dissolve10gBSA(e.g.,Sigma)in100mlH2O.Filtersteril
