
Overview:
ProductName | HSC70ELISAkit |
Description | ColorimetricdetectionofHSC70 |
SpeciesReactivity | Human |
Platform | Microplate |
SampleTypes | Celllysates,Serum,Tissue |
DetectionMethod | ColorimetricAssay |
AssayType | SandwichELISA(Enzyme-linkedImmunosorbentAssay) |
Utility | ELISAkitusedtoquantitateHSC70concentrationinsamples. |
Sensitivity | 1.54ng/ml |
AssayRange | 2.34-150ng/mL |
IncubationTime | 30minutes |
NumberofSamples | 40samplesinduplicate |
OtherResources | KitBooklet,MSDS |
Properties
StorageTemperature | 4ºC | ||||||||||||||||||||||||||||||||||||
ShippingTemperature | BlueIce | ||||||||||||||||||||||||||||||||||||
ProductType | ELISAKits | ||||||||||||||||||||||||||||||||||||
AssayOverview | 1.PrepareStandardandsamplesinStandardandSampleDiluent.2.Add100µLofStandardorsampletoappropriatewells.3.CoverplatewithPlateSealerandincubateatroomtemperature(20-25°C)for1hour.4.Washplatefourtimeswith1XWashBuffer.5.Add100µLofBiotinylatedAntibodyWorkingSolutiontoeachwell.6.CoverplatewithPlateSealerandincubateatroomtemperaturefor1hour.7.Washplatefourtimeswith1XWashBuffer.8.Add100µLofStreptavidin-HRPWorkingSolutiontoeachwell.9.CoverplatewithPlateSealerandincubateatroomtemperaturefor30minutes.10.Washplatefourtimeswith1XWashBuffer.11.Add100µLofTMBSubstratetoeachwell.12.Developtheplateinthedarkatroomtemperaturefor30minutes.13.Stopreactionbyadding100µLofStopSolutiontoeachwell.14.Measureabsorbanceonaplatereaderat450nm. | ||||||||||||||||||||||||||||||||||||
KitOverview |
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CiteThisProduct | HSC70ELISAKit(StressMarqBiosciencesInc.,VictoriaBCCANADA,Catalog#SKT-106) |
BIOLOGicalDescription
AlternativeNames | HSC54ELISAKit,HSC71ELISAKit,HSC73ELISAKit,HSP71ELISAKit,HSP73ELISAKit,HSPA10ELISAKit,HSPA8ELISAKit,LAP1ELISAKit,NIP71ELISAKit |
ResearchAreas | Cancer,HeatShock |
ScientificBackground | HSP70genesencodeabundantheat-inducible70-kDaHSPs(HSP70s).InmosteukaryotesHSP70genesexistaspartofamultigenefamily.Theyarefoundinmostcellularcompartmentsofeukaryotesincludingnuclei,mitochondria,chloroplasts,theendoplasmicreticulumandthecytosol,aswellasinbacteria.Thegenesshowahighdegreeofconservation,havingatleast5O%identity(2).TheN-terminaltwothirdsofHSP70saremoreconservedthantheC-terminalthird.HSP70bindsATPwithhighaffinityandpossessesaweakATPaseactivitywhichcanbestimulatedbybindingtounfoldedproteinsandsyntheticpeptides(3).WhenHSC70(constitutivelyexpressed)presentinmammaliancellswastruncated,ATPbindingactivitywasfoundtoresideinanN-terminalfragmentof44kDawhichlackedpeptidebindingcapacity.PolypeptidebindingabilitythereforeresidedwithintheC-terminalhalf(4).ThestructureofthisATPbindingdomaindisplaysmultiplefeaturesofnucleotidebindingproteins(5).Whencellsaresubjectedtometabolicstress(e.g.,heatshock)amemberoftheHSP70family,HSP70(HSP72),isexpressed;HSP70ishighlyrelatedtoHSC70(>90%sequenceidentity).ConstitutivelyexpressedHSC70rapidlyformsastablecomplexwiththehighlyinducIBLeHSP70incellsfollowingheatshock.TheinteractionofHSC70withHSP70isregulatedbyATP.Thesetwoheatshockproteinsmovetogetherinthecellexperiencingstress.FurThermore,researchonHSC70hasimplicatesitwitharoleinfacilitatingtherecoveryofcentrosomalstructureandfunctionafterheatshock(6). |
References | 1.BrownC.L.,etal.(1993)J.CellBiol.,120(5):1101-1112. 2.BoorsteinW.R.,ZiegelhofferT.,andCraigE.A.(1993)J.Mol.Evol.38(1):1-17. 3.RothmanJ.(1989)Cell59:591-601. 4.DeLuca-Flaherty,etal.(1990)Cell62:875-887. 5.BorkP.,SanderC.,andValenciaA.(1992)Proc.NatlAcad.Sci.USA89:7290-7294. 6.BrownC.L.,etal.(1996)J.Biol.Chem.271(2):833-840. |
ProductImages

TypicalStandardCurvefortheHSC70ELISAkit(Enzyme-LinkedImmunosorbentAssay)StressXpress®–SKT-106.AssayType:SandwichELISA.DetectionMethod:ColorimetricAssay.AssayRange:2.34–150ng/mL.
ProductCitations(1)
OtherCitations
ChangesintheHeatShock70kDaProteinLevelinHumanNeutrophilsInducedbyHeatShock.
Boyko,A.A.,Vetchinin,S.S.,Sapozhnikov,A.M.andKovalenko,E.I.(2014)BioorgKhim.40(5):488-498.
PubMedID:25895348ReactivityHumanApplications:Neutrophil
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pH(1)=pKa+lg[c(CH₃COONa)/c(CH₃COOH)]=pKa=4.74
通HCl后,溶液是c(CH₃COOH)=0.2mol/L、c(NaCl)=0.1mol/L的混合溶液,溶液pH按照弱酸溶液pH的求法求.
c(H⁺)=√[Ka*c(CH₃COOH)]=√(10^-4.74*0.2)=0.00191(mol/L)(采用了近似公式)
pH(2)=-lg{c(H⁺)}=2.72
两个pH求得,那么pH的变化量也就可得了.pH的变化量=|pH(2)-pH(1)|=|2.72-4.74|=2.02
1)PH缓冲溶液作用原理和pH值
当往某些溶液中加入一定量的酸和碱时,有阻碍溶液pH变化的作用,称为缓冲作用,这样的溶液叫做缓冲溶液.弱酸及其盐的混合溶液(如HAc与NaAc),弱碱及其盐的混合溶液(如NH3·H2O与NH4Cl)等都是缓冲溶液.
由弱酸HA及其盐NaA所组成的缓冲溶液对酸的缓冲作用,是由于溶液中存在足够量的碱A-的缘故.当向这种溶液中加入一定量的强酸时,H离子基本上被A-离子消耗:
所以溶液的pH值几乎不变;当加入一定量强碱时,溶液中存在的弱酸HA消耗OH-离子而阻碍pH的变化.
2)PH缓冲溶液的缓冲能力
在缓冲溶液中加入少量强酸或强碱,其溶液pH值变化不大,但若加入酸,碱的量多时,缓冲溶液就失去了它的缓冲作用.这说明它的缓冲能力是有一定限度的.
缓冲溶液的缓冲能力与组成缓冲溶液的组分浓度有关.0.1mol·L-1HAc和0.1mol·L-1NaAc组成的缓冲溶液,比0.01mol·L-1HAc和0.01mol·L-1NaAc的缓冲溶液缓冲能力大.关于这一点通过计算便可证实.但缓冲溶液组分的浓度不能太大,否则,不能忽视离子间的作用.
组成缓冲溶液的两组分的比值不为1∶1时,缓冲作用减小,缓冲能力降低,当c(盐)/c(酸)为1∶1时△pH最小,缓冲能力大.不论对于酸或碱都有较大的缓冲作用.缓冲溶液的pH值可用下式计算:
此时缓冲能力大.缓冲组分的比值离1∶1愈远,缓冲能力愈小,甚至不能起缓冲作用.对于任何缓冲体系,存在有效缓冲范围,这个范围大致在pKaφ(或pKbφ)两侧各一个pH单位之内.
弱酸及其盐(弱酸及其共轭碱)体系pH=pKaφ±1
弱碱及其盐(弱碱及其共轭酸)体系pOH=pKbφ±1
例如HAc的pKaφ为4.76,所以用HAc和NaAc适宜于配制pH为3.76~5.76的缓冲溶液,在这个范围内有较大的缓冲作用.配制pH=4.76的缓冲溶液时缓冲能力最大,此时(c(HAc)/c(NaAc)=1.
3)PH缓冲溶液的配制和应用
为了配制一定pH的缓冲溶液,首先选定一个弱酸,它的pKaφ尽可能接近所需配制的缓冲溶液的pH值,然后计算酸与碱的浓度比,根据此浓度比便可配制所需缓冲溶液.
以上主要以弱酸及其盐组成的缓冲溶液为例说明它的作用原理、pH计算和配制方法.对于弱碱及其盐组成的缓冲溶液可采用相同的方法.
PH缓冲溶液在物质分离和成分分析等方面应用广泛,如鉴定Mg2离子时,可用下面的反应:
白色磷酸铵镁沉淀溶于酸,故反应需在碱性溶液中进行,但碱性太强,可能生成白色Mg(OH)2沉淀,所以反应的pH值需控制在一定范围内,因此利用NH3·H2O和NH4Cl组成的缓冲溶液,保持溶液的pH值条件下,进行上述反应.
这就是说不用酸碱预处理吗?
Whatman的网站上没有DE52最大耐受压力,请问又经验的战友应该是多少?
Whatman的网站上:
DE32DryMicrogranularDEAECellulose
SimilarperformancecharacteristicsafterprecyclingasDE52.
DE52PreswollenMicrogranularDEAECellulose
ProbablythemostwidelyusedDEAEcelluloseintheworld;usedforbiopolymerswithlowtohighnegativecharges;exhibitsexcellentresolutionwithgoodflowrates.
附件是一本图书(MethodsinMolecularMedicine,)的章节,上面说:
WhatmanDEAE52comesalreadypreswollenandonlyneedstobetransferred
totherunningbuffer50mMTE8.
lAntibodiesUsingIonExchangeChromatography.pdf(87.06k)
是否可以理解为纯化水得PH范围为6.3-7.6?能否直接用pH计测量?谢谢!

