| InputType: | HistoneExtracts |
| ResearchArea: | HistoneAcetylation |
| TargetApplication: | AmountQuantitation |
| VesselFormat: | 96-WellPlate |
| 100%Guarantee: | 6months |
The EpiQuik™GlobalAcetylHistoneH3-K9QuantificationKit(Fluorometric) isaconvenientpackageoftoolsthatallowstheexperimentertospecificallymeasureglobalacetylationofhistoneH3-K9fluorometrically, usingavarietyofmammaliancells(human,mouse,etc.)includingfreshandfrozentissues,andculturedadherentandsUSPensioncells. Thekithasthefollowingadvantages:
- Quickandefficientprocedure,whichcanbefinishedwithin2.5hours.
- InnovativefluorometricassaywithouttheneedforrADIoactivity,electrophoresis,orchromatography.
- SpecificallycapturesacetylH3-K9withthedetectionlimitaslowas0.4ng/wellanddetectionrangefrom5ng-2µg/wellofhistoneextracts.
- ThecontrolisconvenientlyincludedforthequantificationoftheamountofacetylH3-K9.
- StripmicroplateformatmakestheassayflexIBLe:manualorhighthroughput.
- Simple,reliable,andconsistentassayconditions.
BackgroundInformation
Acetylationofhistones,includinghistoneH3,hasbeeninvolvedintheregulationofchromatinstructureandrecruitmentoftranscriptionfactorstothegenepromoters.Histoneacetyltransferases(HAT)andhistonedeacetylases(HDACs)playacriticalroleincontrolofhistoneH3acetylationatmultiplesites.HistoneH3atlysine9(H3-K9)isaprimaryacetylatedsiteofhistoneH3.AcetylationofhistoneH3-K9appearstohaveadominantroleinhistonedepositionandchromatinassemblyinsomeorganisms.AcetylationofhistoneH3-K9istightlyinvolvedinthecellcycleregulation,cellproliferation,andapoptosis.AcetylationofhistoneH3-K9isalsoanactiveMarker,andthebalancebetweenhistoneH3-K9acetylationandmethylationisimportantfortheestablishmentofspecificchromatinstructures.AnimbalanceintheequilibriumofhistoneH3acetylation,includingK9acetylation,hasbeenassociatedwithtumOrigenesisandcancerprogression.HistoneH3-K9acetylationmaybeincreasedbyinhibitionofHDACsanddecreasedbyHATinhibition.Thus,quantitativedetectionofglobalacetylhistoneH3-K9wouldprovideusefulinformationforbetterunderstandingepigeneticregulationofgeneactivationandfordevelopingHATorHDAC-targeteddrugs.TheEpiQuik™GlobalAcetylHistoneH3-K9QuantificationKit(Fluorometric)providesatoolformeasuringglobalacetylationofhistoneH3-K9.
Principle&Procedure
Thiskit isdesignedformeasuringglobalhistoneH3-K9acetylation.Inanassaywiththiskit,theacetylhistoneH3atlysine9iscapturedtothestripwellscoatedwithananti-acetylH3-K9antibody.ThecapturedacetylhistoneH3-K9canthenbedetectedwithalabeleddetectionantibodyfollowedbyafluorescentdevelopmentreagent.TheratioofacetylH3-K9isproportionaltotheintensityoffluorescence.TheabsoluteamountofacetylH3-K9canbequantifiedbycomparingtothestandardcontrol.
StartingMaterials
Inputmaterialshouldbepurifiedhistoneextracts.Ingeneral,theinputamountshouldbefrom50ngto200ngperwellofhistoneextracts.
F1(10Xwashbuffer)
F2(antibodybuffer)
F3(detectionantibody,1mg/ml)*
F4(fluorodeveloper)*
F5(fluoroenhancer)*
F6(fluorodilution)
Standardcontrol(100µg/ml)*
8wellsamplestrips(withframe)
8wellstandardcontrolstrips
Userguide
*Formaximumrecoveryoftheproducts,centrifugetheoriginalvialpriortoopeningthecap.
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常用流动相加酸碱后PH的总结，希望大家能够提供一点自己测过的结果，谢谢先
1.直接用固体磷酸钠配制成50mM的磷酸钠溶液，再调pH到7.4；（我们试着用这个做了下，发现挂不上柱）
2.配置磷酸钠盐缓冲液：按NaH2PO4：Na2HPO4以19：81的摩尔比配制成pH7.4的缓冲液？（附一张百度出来的配方
）
3.如果是磷酸钠盐缓冲液，可以直接将50mM的NaH2PO4的水溶液用NaOH调成pH7.4吗？
再者，2和3这两个方法配制的磷酸钠盐缓冲液有什么区别？最终效果是一样的吗？如果不一样，有什么理论的知识支撑呢？个人感觉是分析化学中酸碱理论中的缓冲液那里的知识。求帮忙解答这些疑问。
另外，我还想问一下，pH对于Ni柱对His-tagged的蛋白的分离纯化影响大吗？是怎么影响的？谢谢大家了！
有了源数据之后把源数据按照大小排列，
选中源数据区域-->ALT+A1-->选中图标区右键-->更改图表类型-->散点图
因为是考察不同PH对药物的影响，样品又不好改变其PH值，这种情况怎么办？希望有经验的高手指教。
我的流动相是甲醇-水（90：10）
谢谢赐教！
请进子版按格式发贴，自行修改，谢谢。
由弱酸及其盐、弱碱及其盐组成的混合溶液，能在一定程度上抵消、减轻外加强酸或强碱对溶液酸碱度的影响，从而保持溶液的pH值相对稳定。这种溶液称为缓冲溶液。
:)
我在做一细菌不同酸碱度生长状况时,发现这些奇怪现象:pH=3的培养基灭菌(TSB液体培养基)灭菌后pH上升到到9.2!而原来pH=9.0的降到8.7(基本没多少变化),请问各位大侠,这是什么原因?
一般做不同酸碱度生长实验时,该如何才能防止pH在湿热灭菌后基本不变化?
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!
