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BasicProteinChemistryTechniques
CoomassieBlueStain:(forgels)1)Combine225mlMethanolwith225mlddH2O.2)Add0.5gramsofCoomassieBlue.3)Justbeforeuse,add50mlaceticacidto10%finalconcentration.4)Stainfor2hours.Stainingtimecanbeshortenedbymicrowaving;~50secondsatmediumpower.5)Afterstaining,pouroffthestainintoabottlemarkedUsedStain.Staincanbe...
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整肠生治疗80例抗生素相关性腹泻的临床观察
...(pdf)【摘要】目的观察整肠生胶囊防治抗生素相关性腹泻(ADD)的作用。方法使用整肠生胶囊来治疗80例ADD。结果通过随机、双盲、对照试验证实整肠生组显效率达82.14％，总有效率达95％左右。因为即使出现了ADD还可以继续使用抗...
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Radioiodinationofprotein
...ation(byJunTakagi,6/16/2000)PurposeandbackgroundsPrincipleofradioiodinationAdditionofoxidizingreagents(suchaschloramine-Torperoxidase+H202)convertsI-toI+orI3-.Thishighlyreactivemoleculeattackso-positionoftyrosine(orinsomecase,histidineisalsolabeled).Aboutnuclide...125-Idecay:electroncapture(g-ray,0....
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DGKAssay
...-Dyerextraction:a)Resuspendcellpelletin3mlchloroform/methanol(1:2)vortex.b)Add0.8mlwatervortex.c)Pelletcellulardebrisbyspinningat3,000rpm(2000xg)for5min.d)Add1mlchloroformvortex.e)Add1mlwatervortex.f)Discardupper,aqueousphase.3)Collect1.2-1.5mlofthelowerphase(totalvolumeoflowerphaseshouldconstitute2...
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常用试剂配制－3
...ed.Aceticacid(MW60.05)Glacialaceticacidis99.6%(w/v)aceticacid,andis17.4M.1MAdd57.5mlofglacialaceticacidto800mlofwaterandthenmaketo1literwithwater.0.05NAdd2.87mlofglacialaceticacidto800mlofwaterandthenmaketo1literwithwater.0.9MAdd54mlofglacialaceticacidto800mlofwaterandthenmaketo1literwithwater.7%(w/...
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SphingomyelinQuantitationPost-cholineLabelingofHL60Cells
...urbthepelletsinceitissomewhatdispersed.4)Immediatelyuponaspiratingeachtube,add3mlsofchloroform-methanol,1:2,andvortexvigorously!!--ItisnecessarytoaddtheC:Mimmediatelyinordertogetgooddispersionofthepellet.Iftoomuchtimepassespriortoresuspension,thepelletdoesnotdissolvewellandsonicationisnecessary.--Th...
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SphingomyelinMassMeasurement
...roximately1X107cells.2)Resuspendpelletin3mlCHCl3:CH3OH(1:2)andvortexhard.3)Add0.8mlH2O,vortexhardandallowsolutiontositatroomtemperatureforatleast30minutes.4)Pelletcellulardebrisbyspinningat3,000rpm(inthetabletopcentrifuge)for5minutes.5)Transfersolventstoanewtubewithouttransferringanyofthepelleteddeb...
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SpotFluorescence法定量DNA
...r(ageltrayisfine).2.Spotseveral3μldropsofTEcontaining2mg/mlEtBrontray.3.Add1μlofaseriesofknownDNAconcentration(lDNAat0,1,2.5,5,1020ng/μl)standardstotheEtBrdrops.Mixbypipettingupanddownseveraltimes.4.Add3dilutions(1/2,1/4,1/8)ofeachofyourunknownDNAsamplestotheethidiumbromidedropsasfollows:Mi...
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AA&MetaboliteQuantitationofCellPelletsPost-AALabeling
...inceitissomewhatdispersed.2)Immediatelyuponremovingsupernatantfromeachtube,add3mlsofchloroform-methanol,1:2,andvortexvigorously!!--ItisnecessarytoaddtheC:Mimmediatelyinordertogetgooddispersionofthepellet.Iftoomuchtimepassespriortoresuspension,thepelletdoesnotdissolvewellandsonicationisnecessary.3)Ad...
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InvitroSphingomyelinaseAssay
...hevolumeofthismixshouldbe100µl.13)Pre-incubatefor10minutesat37°C.14)Add100µl14C-SMmixtoeachsampleandmixgently.15)Incubatereactionfor15minutes.16)Stopreactionsbyadding1.5mlofchloroform:methanol,1:2.17)Vortex,add0.2mlofwaterandvortex.18)Add0.5mlchloroformandvortextobreakphases.19)Add0.5ml...
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PreparingaSelenomethionylProtein
...c.),and150uLofdayculture.Growat37°Covernight.Day2：ToeachliterM9(1xconc.)add:10mL20%Glucose(sterilefilteredorautoclaved)2mL1MMgSO4(sterilefiltered)0.05mL2MCaCl2(sterilefiltered)0.1mL0.5%(w/v)thiaminesolution(sterilefiltered)1mLantibiotics(1000xconc.)20mLaminoacid50xStock(Ifprecipitateisseen,heatto...
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传导性角膜成形术治疗老视医学验光的体会
...调节幅度。装上近用测试卡在33cm的距离。双眼同时引入Add测量程序，令患者注视视标并比较横线和竖线的清晰度，如果横线清晰就在双眼前同时增加正球镜度数，竖线清晰则降低正球镜度数。直到横竖线一样清晰为止，此时的...
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传导性角膜成形术治疗老视医学验光的体会
...调节幅度。装上近用测试卡在33cm的距离。双眼同时引入Add测量程序，令患者注视视标并比较横线和竖线的清晰度，如果横线清晰就在双眼前同时增加正球镜度数，竖线清晰则降低正球镜度数。直到横竖线一样清晰为止，此时的...
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与超级细菌赛跑：寻找新型抗生素
...大学分子生物研究所领导的开放式抗菌药物发现组织（CO-ADD），发起了“全球搜寻新抗生素项目，邀请全球化学家提交自己的化合物，进行抗菌活性筛查。CO-ADD发言人马克·布莱斯科维奇称，未来具有高耐药性的细菌很可...
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溴乙非啶的处理
...esthemutagenicactivityby~3000foldandistakenfromQuillardetandHofnung,1988.1)Addsufficientwatertomaketheconcentration0.5mg/ml.2)Add1volumeof0.5MKMnO4followedbyonevolumeof2.5MHCl.Mixcarefullyandallowtostandatroomtemperatureforseveralhours.N.B.:AcidifiedsolutionsofKMnO4arepowerfuloxidisingagents.3)Add1v...
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LOWRYPROTEINASSAY
...thprotein.Whenfolinphenolreagent(phospho-molybdic-phosphotungsticreagent)isadded,theFolin-phenolreagentbindstotheprotein.Boundreagentisslowlyreducedandchangescolorfromyellowtoblue.Whilewidelyused,theLowryprocedureislesspreferableanassaythansomeotherproteinassayssinceitismoresubjecttointerferencebyaw...
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Phosphoaminoacidanalysisofnon-TCAprecipitableproteins
....Cutoutgelslice,incubate60minwith1ml5.7MHCl@110oCin13x100mmscrewcaptubes.2.Add4mlwater,centrifuge@2000rpminbenchtopcentrifugefor10min,transfersupernatant,freezeandlyophilise.3.Dissolveresiduein10mlwater,add0.4ml50%(v/v)slurryofDowexAG1X8(Cl-form),adjustpHto7.5-8.5withNH4OH.4.Add1µlofsolutionco...
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AA&MetaboliteQuantitationofMediaPost-AALabeling
...ember4,19941)Remove2500µlaliquotsofsupernatantintoscintillationvials,addscintillationfluidandcount.2)Aliquot1.6mlsoftheremainingsupernatantintoeachoftwopyrexglasstubes.3)Add6mlsofchloroform-methanol,1:2,andvortexvigorously.--Stillhaveamonophaseanditmaybebesttoallowthismonophasetoequilibratefor...
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Lipidanalysis：EXTRACTIONOFLIPIDSFROMLIPOPHORIN
...poproteinsareexposedtothesolventmolecules.Subsequently,waterandchloroformisaddedtoshifttherationto2:2:1.8,nowthepolarandnonpolarsolventsarenolongermiscible,andwegettwophases.Note:Chloroformisheavierthanwater.Thespeedandeaseofthismethodmakeitsuitableformultipleextractions,e.g.ofplasmasamples,provided...
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PhosphateAssaybySupryaJaydev
...,5,7,10,12,and15µlofa1mMNaH2PO4stocksolution2)Aliquotlipidunknowns.3)Add100µlashingbuffertoeachsample.4)DryEtOHandchloroformfromsamplesbyheating@80°Cfor~20minutes.5)Flamesamplesusingastrongflameonthebunsenburner.6)Removesamplesfromflamewhenthebrowngasceasestobereleased.--Thereshouldbeaw...
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高效液相色谱法测定萘丁美酮的含量
...［4］，也有气相色谱法［5］和重量法［6］的报道。BP1993Add1996开始收载萘丁美酮质量标准，采用高效液相色谱法外标法测定萘丁美酮的含量及有关杂质；本文所述高效液相色谱法，分离效果好而且准确，结果令人满意。1 仪器...
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Bradfordproteinassay
...incomplexsolutionisconstantovera10-foldconcentrationrange.：Equipment：Inadditiontostandardliquidhandlingsuppliesavisiblelightspectrophotometerisneeded,withmaximumtransmissionintheregionof595nm,ontheborderofthevisiblespectrum(nospeciallamporfilterusuallyneeded).Glassorpolystyrene(cheap)cuvettesmay...
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高效液相色谱法测定萘丁美酮的含量
...［4］，也有气相色谱法［5］和重量法［6］的报道。BP1993Add1996开始收载萘丁美酮质量标准，采用高效液相色谱法外标法测定萘丁美酮的含量及有关杂质；本文所述高效液相色谱法，分离效果好而且准确，结果令人满意。1 仪器...
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新技术：In-CellWesternAssay
...edsecondaryantibodies(MolecularProbes,Inc.)OdysseyBlockingBuffer(927-40000)AdditionalReagents：1XPBSwashbufferTissueculturereagents(serum,D-MEM,trypsin,1XPBS)20%Tween-20EpidermalGrowthFactor(UpstateGroupInc.,01-107)37%formaldehyde10%TritonX-100Nunc™96Microwell™Plate(NuncPartNumber167008...
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CT图像减影法的临床应用价值初探
...螺旋CT机，操作系统版本号为HISPEEDSERIES6.03，其上装载了ADD/SUB（+/-）功能，对ADD/SUB的临床应用，尚未见报道，笔者通过临床实践，认为减影法有一定的临床应用价值，现报告如下。 1方法 在GE公司CTHISPEEDSERIES6.03操作系统...
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cAMP分析cAMPAssays
...ISAplateBoilingwaterbathorheatingblock,presetto100°CIncubatorpresetto30°CAdditionalEquipmentandReagentsThisprocedurealsorequiresequipmentfordialysisandforthepreparationofcAMP-BSAconjugate(seestep1fordetails).METHODPreparecAMP-BSAconjugateasfollows(modifiedfromJosephandGuesdon1982).Prepareasolution...
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房颤治疗的困惑与希望
...TAF研究者认为“节律控制未能体现出优越性的原因是由于ADD长期疗效难以保证”。分析原因可能与以下几方面因素有关：首先是通过ADD来维持窦性心律的效果有限。CTAF研究发现服用胺碘酮、索它洛尔及普罗帕酮后随访至16个月时...
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Lowry–ProteinDetermination
...50Prot.conc.,ug/ml010205010020050010002000Method1.To0.2mLofsampleorstandardadd1mLoffreshlymixedcomplex-formingreagent.Letthesolutionstandatroomtemperaturefor10min2.Add0.1mLofdilutedFolinreagent,usingavortexmixer,andletthemixturestandatroomtemperaturefor30-60min(donotexceed60min)3.Readtheabsorbanceat...
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SouthernBlots技术
... 20%SDS(OR10%SDS)0.3(OR0.6) Total30mlfor2blots.65℃预热. AddpreparedssDNA（10mg/L）≥0.3ml 倒入杂交液，加入尼龙膜，65℃预杂交4~5小时（新膜） 注意：赶尽膜与膜之间的气泡后把几张膜同时放入杂交管，并用玻棒赶尽膜与管...
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荧光测DNA浓度
...eotherwillreceivenodye(theblank).TheDNAcanbedilutedinTE,butthemaximumvolumeaddedtotheassaytubesshouldbe5microlitersorless.3.AliquotamountsoftheunknownDNAtotubesintherangeofthestandardcurve(preferablyinthemidpointoftherange).4.Add200microlitersofPanVeraDNAassaybuffertoeachtubeandmixwell(Important:the...