MTT cell proliferation assay kit, 1000 reactions. Kit content: Solution A 10ml, Solution B 100ml. MTT Proliferation Assay Kit provides an easy to use tool for studying the induction and inhibition of cell proliferation in any in vitro model. This kit will also allow investigators to screen drug candidates involved in cell cycle regulation. In this assay, MTT is taken up by cells through the plasma membrane potential and then reduced to formazan by intracellular NAD(P)H-oxidoreductases
MTT Cell Proliferation Assay Kit
Catalog Number:
CP-001, 500 reactions
CP-002, 1000 reactions
Introduction
The reduction of tetrazolium salts is now recognized as a safe, accurate alternative to radiometric testing.MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay, first described by Mosmann in 1983, is based on the ability of a mitochondrial dehydrogenase enzyme from viable cells to cleave the tetrazolium rings of the pale yellow MTT and form a dark blue formazan crystals which is largely impermeable to cell membranes, thus resulting in its accumulation within healthy cells. Solubilization of the cells by the addition of a detergent results in the liberation of the crystals which are solubilized.The number of surviving cells is directly proportional to the level of the formazan product created. The color can then be quantified using a simple colorimetric assay. The results can be read on a multi-well scanning spectrophotometer (ELISA reader).
Storage:
Upon receiving, Solution AMTT solution should be kept at-200C and protected from light.Store properly, the kit components should remain stable for 6 months.
Kit Contents:
1.Solution AMTT solution: 5ml ( 500 reactions), 10ml (1000 reactions)
2. Solution BCrystal dissolving solutions: 50ml ( 500 reactions), 100ml (1000 reactions)
Protocol
1. Plate cells in 96-well tissue culture plate at density of 2x105 per well in 100ul of culture medium.
2. 5 hours before the end of the incubation add 10ul of MTT solution from step one to each well containing cells.
3. Incubate the plate at 37ºC for 5 hours.
4. Remove media with needle and syringe.
5. Add 100ul of Solution B to each well and pipette up and down to dissolve crystals.
6. Transfer to plate reader and measure absorbance at 570nm.
Reference Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods. 1983 Dec 16;65(1-2):55-63.
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1970科家致癌RNA病毒发现种特殊RNA病毒聚合酶,该酶能RNA模板,根据碱基互补原则.程与般病毒转录向相反,故称逆转录,催化程酶称逆转录酶,发现哺乳物胚胎细胞裂淋巴细胞含.
逆转录病毒称携带逆转录酶病毒,先侵入宿主细胞病毒RNA模板,靠酶形DNA环化,合宿主细胞染色体原病毒形式宿主细胞代代传.
HIV（艾滋病病毒）典型逆转录病毒
我是新手。我要开始做实验了，但一开始就碰到了个很大的问题。
我想买个能利用哺乳动物细胞RNA聚合酶将DNA转录成RNA的的质粒，向好几家公司打了电话，但都没有我想要的东西。
没有这个东西，我的实验就只能是蓝图了。哪位战友有办法的，能不能帮帮我？
不胜感激。
ResponseofskeletalmuscleRNApolymerasesIandIItotumourgrowth
BiochimicaetBiophysicaActa(BBA)-GeneStructureandExpression,Volume950,Issue3,7September1988,Pages296-302
GeorgeA.J.Goodlad,CatherineM.Clark
或者有高手知道怎么测也行
最近遇到了实验上的大问题，就是用一步法做RNA的荧光定量PCR时，发现低浓度的做不出来，而DNA是可以的，想请问各位在一步法PCR中有没有什么高见，就是逆转录的Buffer有没有什么需要注意的地方！就是在逆转录酶的最适Buffer和Taq酶的最适Buffer中寻找一个最适的Buffer！
