ProductDescription
RibonucleaseHfromE.coli
RibonucleaseHisanendonucleasewhichspecificallydegradestheRNAmoietyofaDNA:RNAhybrid,withoutaffectingtheunhybridizedRNA.Itdoesnotaffectdouble-strandedRNAorDNA.Itisprovidedina50%glycerolstoragebuffer.ItisalsoavailableinaTrehaloseBuffer(Suitableforlyophilization).Concentration1000U/mL.
Pleaseinquireforcustomconcentrationsandbulkquantities.
Applications:
Enablingthesynthesisofsecond-strandCDNAbyremovaloftheRNA
UsedinconjunctionwithAMVRTandT7RNAPolymeraseinamplificationofRNA
UnitDefinition:
OneunitofRNaseHisthatamountofenzymewhichcatalysistheproductionofonenmolofacid-solublenucleotidein20minutesat37ºCusingthefollowingreactionconditions:
40mMTrisHCl,pH7.5
1.0µM[3H]-poly(rA):24µMpoly(dT)
4.0mMMgCl2
1.0mMDTT
30µg/mLBSA
4.0%glycerol
StorageBufferConditionsofRNaseHinGlycerol:
20:0mMTrisHCl,pH7.5
300mMKCl
20.0mMMagnesiumAcetate
7.0mMEDTA
1.0mMDithiothreitol
50%Glycerol
0.2%TritonX100
StorageBufferConditionsofRNaseHinTrehalose.Sameasabove,exceptfor1.0MTrehaloseinsteadofGlycerol.
QualityControl:
DNaseActivity:
One-halfµgofHaefragmentsofPhiX-174DNAisincubatedat37ºCwith2.5unitsofRNaseHfor3hours,andthenelectrophoresedinanativeagarosegelsimultaneouslywithcontrolpositiveDNase1reactions.Nomorethantheequivalentof2.5X10E-4unitofDNase1isdetected.
RibonucleaseActivity:
OnemicrogramofanRNALadderisincubatedfor2hoursat37ºCwith4.0unitsofRNaseH,andthenelectrophoresedinanativeagarosegelsimultaneouslywithcontrolpositiveRNase1Areactions.Nomorethantheequivalentof8X10-8unitofRNase1Aisdetected.
SpecificActivity:
ThespecificactivityoftheE.coliRNaseHisnolessthan300,000unitspermg.
References:
Sambrook,J.,Fritsch,E.F.,andManiatis,T.(1989)MolecularCloning:ALaboratoryManual,(2ndEd.),8.64–8.65
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请教各位老师,
文献中检测细胞的活性有的用“NADPHdehydrogenase(NADPH脱氢酶)”有的用“NADPHdiaphorase(NADPH黄递酶)”
它们是同一种酶吗?它们的功能是什么?检测它们的活性是否能够评估细胞的活性。
不胜感激
如果能分解,那小肠液中的消化酶如何大量共存,如果不能
那它如何识别其他蛋白质物质是不是消化酶?
反应体系如下:
plasmid10ul
10xKbuffer5ul
KpnI1ul
BamHI1ul注意千万不可多加总酶量必须<4%
ddH20upto50ul
总体积改变加酶量按比例改变.
明白了么,少年?
有谁用碧云天的过氧化氢酶检测试剂盒,在试剂盒的准备工作中,过氧化氢的实际浓度=22.94*A240,我测出来的吸光度是3.3左右,那么乘以22.94就等于76多点,再乘以之前的稀释倍数,大约就是7600mM左右,这样跟说明书中说道的1M相差太多,感觉不对啊,稀释的肯定是没有错的,但是不知道哪里出了错,怀疑说明书就有问题呢,有人做过这个实验吗?能不能准确测出过氧化氢浓度吗?有测过的人帮忙指点一下啊,不知道应该怎么做