Introduction Over one third of diabetics will develop diabetic kidney disease (DKD) in their lifetime1. Despite the large amount of diabetics suffering from DKD, current methods for measuring and tracking kidney function may not be specific enough for diabetic systems2,3. […]
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Catalog #54-043SpeciesCanineRegulatory StatusResearch Use Only. Not for Use in Diagnostic Procedures.Product DistributionAvailable in North America OnlyRange4 - 400 pg/mLSensitivity0.56 pg/mLSizes96 WellsSample TypesCell Culture, Plasma, Serum, Tissue Culture, UrineIncubation Time3 hrIncubation Time3 hr 10 minSample Size10µLInstructions for UseClick here to viewSafety Data SheetClick here to view - Additional Information
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NGAL (neutrophil gelatinase-associated lipocalin) belongs to the lipocalin family of proteins. These are small secreted proteins characterized by their ability i) to bind small hydrophobic molecules in a structurally conserved pocket formed by β-pleated sheet, ii) to bind to specific cell-surface receptors, and iii) to form macromolecular complexes. NGAL has many synonyms: perhaps the most widely used is lipocalin2 (LCN 2); more recently the name siderocalin has been used to express NGAL’s ability to bind bacterial siderophores. As NGAL was first reported as oncogene 24p3 in the mouse, this name is still occasionally used to identify NGAL in other species.Because dog NGAL has not yet been isolated and no antibodies or immunochemical detection methods have hitherto been developed to detect or measure it, there are no published data on the physiological or pathological functions of this protein in the dog. However, it is likely that some features of the biosynthesis, location and release of dog NGAL will be similar to those of its homologues in other species. The present kit has in fact been developed to extend NGAL research and the use of NGAL as a marker molecule, especially for kidney injury, to the dog. In the following, some pathophysiological features of NGAL in other mammalian species will be mentioned, because similar features may be observed in the dog.Like other mammalian NGALs, dog NGAL may in certain situations be co-expressed and form complexes with matrix metalloproteinase-9 (MMP-9). Dog NGAL may also prove to form homodimers and higher oligomers with itself. As dog NGAL, unlike its human homologue but like all other mammalian NGALs known, does not possess a third cysteinyl residue capable of forming an intermolecular disulfide bridge, any complex formation detected will probably depend on non-covalent forces, at least initially.
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DNA连接酶作用于基因工程,用于连接两个DNA片段间的磷酸二酯键。
但是也有以RNA为模板的DNA聚合酶和RNA聚合酶 (也就是反转录酶和RNA依赖的RNA聚合酶),它们的结合位点在RNA上.
也就是说,模版是谁,结合位点就在谁上。
2.结合位点就是启动合成DNA或者RNA的碱基序列,如RNA聚合酶结合位点是转录起始位点,是一段特殊的位于编码基因上游的DNA序列.这段DNA序列可以结合RNA聚合酶,从而起始转录过程.经典的RNA聚合酶结合位点是TATA box.
TATA box是编码序列前的4个碱基.在大多数生物的基因前面都有TATA着4个碱基序列的存在,它就是一个RNA聚合酶结合位点.
最近,我看了很多这方面的资料,但是我对这个方面不是很了解,所以现在搞得头昏眼花的,不知道该选什么酶和载体,希望大家多多帮忙
在此十分感谢
再来说真核生物,真核生物的DNA聚合酶分为α,β,γ,δ,ε,.首先我来说三个和原核生物中的Ⅰ,Ⅱ,Ⅲ功能相同的酶.ε类似于原核生物DNA聚合酶Ⅰ,β类似于原核生物DNA聚合酶Ⅱ,δ类似于原核生物DNA聚合酶Ⅲ.α具有引物酶活性,而γ则是作为复制真核生物线粒体内的DNA所需要的酶.
下面是六种不同耐热聚合酶的比较:
Taq:扩增效率最高的耐热DNA聚合酶,能很好的扩增6kb以下的DNA片段。扩增碱基出错率为10-5左右。
Pfu:目前保真度最高的耐热DNA聚合酶,碱基出错率为10-6,但扩增效率低于Taq酶,一般能很好的扩增2kb以下的片段。
TaqPlus:集扩增效率高和保真度好于一身。扩增效率比Pfu高,保真度比Taq好。能有效的扩增10kb以下的片段。
HotstartTaq:经过化学修饰的耐热DNA聚合酶。此酶在常温下,活性被化学基团封闭,要在94℃-95℃加热数分钟才能回复正常活力开始反应,避免了起始循环较低温度下的非特异性扩增,提高了反应的灵敏度和特异性。
LongTaq:具有3’-5’外切酶活性的耐热DNA聚合酶,它不但扩增效率高而且错配率低,对于简单模板可扩增长达40kb的模板,对复杂模板也可扩增长达15kb的片段。
TaqPlatinum:热启动高保真耐热DNA聚合酶。如果对保真度要求很高,而用Pfu扩增有难度,可选用TaqPlatinum,一般扩增长度可达4kb。
根据不同的实验目的选择最合适的酶,
克隆普通长度的目的DNA片段:Taq、TaqPlus
保真度要求较高,片段比较短,如点突变、基因筛选等:Pfu、TaqPlatinum
高保真长片段扩增,如构建基因图谱及分子遗传学研究等:LongTaq、TaqPlus
扩增基因组模板,需要降低背景:HotstartTaq、TaqPlatinum
扩增GC含量较高或二级结构较复杂的模板:TaqPlus、LongTaq、TaqPlatinum
实时荧光定量PCR反应:Taq、HotstartTaq、TaqPlatinum
从菌株或质粒模板,筛选鉴定目的克隆,扩增6kb以下的片段:Taq、TaqPlus、2×TaqPCRMasterMix
模板比较复杂或目的片段丰度低,用普通Taq酶扩增不出条带:2×TaqPCRMasterMix、2×PfuPCRMasterMix
我用来扩增一个1.1kb的基因,用TAKARA的EXtaq和其他的总是出现突变不知道这个酶能不能完全保真,另外末端是否加A
聚合酶就是多功能的dna的制作机器


