Hi,

Try the BrDU kit by Millipore(earlier used to be from Calbiochem).Cat no.QIA58.

Regards

Charan

I agree to Holger Fey. We tried a lot of different kits in our lab, but each had disadvantages and results were inconsistent. We now just  count cells using a FACS.

Best regards Jörg

I'll also second Holger Fey. Good old haemocytometer with trypan blue or if you have access to a Coulter counter is the easiest and most reproducible way of measuring cell number. All the assay describe by Ilias are dependent on metabolic activity of the cells and in some conditions might not represent the actual cell number. For example, a confluent dish of cells might show a lower number of cell than a subconfluent actively growing dish of cells by MTT/Alamar blue just because of lower metabolic activity.   

For cell proliferation, we use the Beckam Coulter Vicell to good effect which counts cell number and viability. Downsides are that you have to have a minimum volume of 500µL and minimum of around 10,000 cells for the count to be accurate. 

I have tried a lot of different kits in our lab. At the begining I would start with SRB test which is frequently compared to MTT, but is much cheaper. MTT assay requires cellular methabolic activity, whereas SRB does not distinguish between viable and dead cells. However, this diffrence does not compromise the ability of  the SRB assay to detect cytotoxic effects.

Teh problem with all these kit is, that you just assume to have cell growht. BrdU incorporation is not neccessarily meaning cells are proliferating (somme cell types can produce several nuclei). Same with all these mitochimdria bases assays (MTT...) So I go with the majority and would recommend counting. However if you know that cellsare proliferating you can use DNA incorporationassays or all these other kits but tryphan blue counting ist easy cost effective and precise

Regards

Sven

I've had really good luck with the invitrogen Click-iT EdU kit. I've used it on MDA-MB 231 cells in culture, it works great and it's easy to use.

I second the recommendation from Santiago -the Click-it EdU kit works really really well. A bit pricy, but I think it's worth it because its a really easy protocol and the results are very clear.  

I have had very good results with CyQuant assay.

you can use Click-iT EdU Alexa Fluor 488 imaging kit (C10083)

My favourite proliferation assay which I have used over the years would have to be Cell Titer Blue from Promega. This assay is very easy to used, non toxic to the cells and can be used as a fluorescence assay which is really sensitive but if you do not have the correct filters in your flourimeter it can also be used as a colorimetric assay. 

This is a little bit of manuel method but CFSE assay really good for proliferation detection... if your lab hava flow cytometry you can easilly use this method 

I recommend Dojindo or Promega.

I recommend MTS from promega. it is really good and allows you to work with the 96 wells plate and test many condition all toghether. good luck

https://www.promega.es/products/cell-health-and-metabolism/cell-viability-assays/celltiter-96-aqueous-one-solution-cell-proliferation-assay-_mts_/

I worked with CyQuant NF cell proliferation assay kit from Life technologies and it works good. 

You can use cell proliferation reagent WST-1 from Roche. The catalong no is 11 644 807 001

Brdu Assay Kit (SIGMA)

Why don't you take the Buerker chamber or Coulter counter and count the cells over time???

I use CyQUANT (linked below).  It's relatively easy to use (the trickiest part is getting your initial cell counts perfect), it's cheap (about $200 for 1000 assays), and it's independent of metabolic activity (as opposed to something like an MTT assay). 

http://www.lifetechnologies.com/order/catalog/product/C7026

For Non-Radioactive proliferation assay, MTT reagent assay is quite convinent and reliable to use. If you perform assay using this kit, Duplicate the experiment for confirmation the accuracy of the experiment. Thank you

https://ita.promega.com/products/cell-health-and-metabolism/cell-viability-assays/celltiter-96-non_radioactive-cell-proliferation-assay-_mtt_/?activeTab=0

I'm using the MTS assay by promega... shorter time compared to the MTT, comparable price, cells after the assay can be used for other assays because the protocol is non-lytic.

Promega also have different assays based on ATP, DNA and other biological target (you can also use LDH assays lysing cells after treatment... it's an indirect measurement)

We are using Cell Counting Kit - 8 from Sigma. Cell Counting Kit-8 allows sensitive colorimetric assays for the determination of the number of viable cells in the proliferation and cytotoxicity assays. The detection sensitivity is higher than any other tetrazolium salts such as MTT, XTT or MTS.

Tetrazolium salt assays (MTT, XTT) are about as simple and easy as they get. life tech, signma aldrich, and ATCC all sell kits. all you need is a well plate  and plate reader with the appropriate filter. 

Thank you everyone for your suggestions. In our lab we discuss this issues quite a lot and MTT/XTT these assay usually define the metabolism of the cells and not actual proliferation. Whereas on the other hand, BrDU incorporation gives us the right information when it incorporates into the DNA. THus when cells are actually proliferating, we come to know how much of it is proliferating compared to control.

Is mitochondria a good marker for proliferation assay? Based on treatment conditions, mitochondria will signal in two directions either positive or negative for the nuclear replication or cell cycle progression. We need to consider both of them and also need to think about quality vs. quantity. This is my concern.