MTT细胞增殖检测试剂盒详细说明

 
   
MTT细胞增殖检测试剂盒详细说明MTT Cell Proliferation Kit 
Package: 500 Tests 
Description 
Cell proliferation assays have been widely used to assess cell cycle regulatory factors such as growth factors, cytokines, 
mitogens,  and  drugs.  Tetrazolium  salts  (e.g.  MTT,  XTT,  WST-1  and  WST-8)  are  especially  useful  for  assaying  the 
quantification of viable cells, because they are cleaved to form a formazan dye only by metabolic active cells. 
MTT Cell Proliferation Assay Kit provides a convenient tool for studying the induction and inhibition of cell proliferation in 
vitro. In this assay, MTT is taken up by cells due to its net positive charge and the plasma membrane potential. It is reduced 
to formazan by intracellular NAD(P)H-oxidoreductases. This kit also allow investigators to screen drug candidates involved 
in regulation of cell cycle. 
Kit Components 
Component  250 Tests  500 Tests 
Component A: MTT Reagent 
Component B: MTT Solvent 
Component C: Formazan Solubilization Solution 
12.5 mg X 1 
1.25 mL X 2 
27.5 mL X 1 
25 mg X 1 
5 mL X 1 
27.5 mL X 2 
Protocol  1  1 
Storage 
The MTT powder and MTT Solvent should be stored at 2-8°C with protection  from  light. Formazan Solution should be 
stored at room temperature. 
Procedure 
Note: 
Thaw the kit component at room temperature before use. 
1.  Centrifuge before opening all the tubes. 
2.  Seed cells at a density of 2000 cells/well for cell proliferation assay and 5000 cells/well for cell cytotoxicity assay in 
100 μL cell culture medium in 96-well plate. Incubate the plate for 24 hours in a humidified incubator. 
Note: 
5 repeated well for one sample is recommended for each assay. 
3.  Dissolve MTT Powder (Component A) with MTT Solvent (Component B) to prepare MTT Reagent (5 mg/mL). 
4.  Add 10 μL of MTT Reagent (prepared above) at a final concentration of 0.5 mg/mL to each well. 
5.  Mix gently and incubate for 4 hours in a CO2 incubator. 
Note: 
The formazan produced in the cells will appear as dark crystals in the bottom of the wells after incubation. 
6.  Aspirate the culture medium from each well carefully to prevent disruption of the cell monolayer. 
7.  Add 100 μL of Formazan Solubilization Solution (Component C) into each well. 
8.  Put the plate on a shaker to mix gently for 10 minutes to dissolve the formazan crystals. 
9.  Measure the absorbance at 570 nm using a microplate reader. 

   
Before use, please read this protocol carefully. For your safety, please use gloves, goggles or other protective measures.   
This kit is for R&D use only, not for drug, household or other uses.