Details
Applications
- Preferedsubstratesaresingle-strandedDNA,double-strandedDNAwith3’-hydroxylterminiandoligodeoxynucleotideprimers(1)
- Usedforspecificlabelingof3"-terminiwithribonucleotides(2)
- Labels3"-endsofDNAfragmentswithan[α-32P]3"-deoxynucleoside(3)
- AddshomopolymertailsofdeoxyribonucleotidestovectorsorCDNAs(4,5)
ReagentsSupplied
5XTdtC-TailingReactionBuffer
UnitDefinition
Oneunitistheamountofenzymerequiredtotransfer1nmolofdAMPfromdATPtothe3"OHterminusoftheoligonucleotideinitiatord(A)15for1hourat37ºC
HeatInactivation
15minutesat 75ºC
Concentration
10-15units/µl
StorageBuffer
50mMpotassiumphosphate(pH7.4)
1mMβ-mercaptoethanol
50%(w/v)glycerol
AssayConditions
40mMKCacodylate(pH7.2)
8mMMgCl2
0.33mMZnSO4
10µMoligonucleotided(A)50
10µl bovineserumalbuminbuffer
1mM(α-32P)dATP
Reactionvolumeof60µl
Note:Allreactionsshouldberuninpolypropylenetubes
StorageConditions
Storeat-20°C
ProductshippedonDryIce
Downloads
CertificateofAnalysisPDF -CurrentLot
MSDSPDF -CurrentLot
References
(1)Chang,L.M.andBollum,F.J.(1971)J.Biol.Chem.246,909-916
(2)Roychoudury,R.andKossel,H.(1971)Eur.J.Biochem.22.310-320
(3)Tu,C.P.D.andCohen,S.N.(1980)Gene10,177-183
(4)Roychoudhury,R.,Jay,E.andWu,R.(1976)NucleicAcidsRes.3,863-877
(5)Deng,G.andWu,R.(1983)MethodsEnzymol.100,96-116
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这些染料都非常成熟,光毒和淬灭都很低,当然要考虑到你采集图像时的显微镜参数。
比如calcein,常用的示踪,绿光(虽然这些颜色只是根据光谱加上去的伪彩),但要考虑你的实验过程中结合细胞结构,是否会伴随calcein的泄漏,就是荧光降低。
dri,还可以看膜啊
cfda也不错
bcef虽是ph指示,但你试验时不仅可观察细胞,还可看细胞ph变化,也行。
当然你或许还要结合其他方法,如细胞免疫化学等手段去双染或多染,都需要综合考虑染料之间特性。单独染一个染料,有点浪费,不如多染,数据和图像也好看些。现在流行细胞成像。

