Question

Can you please write about the advantages and disadvantages and some troubleshooting of WST-8 assay, BrdU assay, and a clonogenic assay. Thank you in advance. I would appreciate it a lot!!

Answer 1

WST-8 :

These are the assay that are used to determine the dehydrogenasing  activities in the samples and also used for screening the substrates. It is also used to perform various assay such as cell viability, cell proliferation assay etc.

Advantages :

• These have very low cytotoxicity , because it is not a cell permeable
• It requires simple assay procedures and does not requires any extra steps to dissolve.
• Cellular reduction occurs and produces an water soluble formazan.
• It is also used for quantitative measurement of dehydrogenase activities.

Disadvantages :

• Changes in the intracellular metabolic activities results in reduction of assay substrates .
• It involves toxicity

BrdU assay  :

It is an chemical structure which is used to determine cell proliferation assays and it is also used replicating cells. cell proliferation are done in wide variety of species from plants to animal cells.

Advantages :

• It is very stably integrated with DNA ,helps in measuring cells for months.
• Maintains the cell integrity
• Assay requires less time i.e low time consumption
• Main advantage is it constitutes a non radioactive technique.

Disadvantages:

• It involves in the harsh treatments of tissue sections
• Technically it is  cumbersome
• The difficulty can result in tissue damages
• No frequent assays can be performed with a cell

Clonogenic assay :

Clonogenic assay is nothing but an cell biology technique used for study the survival of the irradiated cancer cells. Cell survival assay are based on the ability of a single cell to grow into a colony.

Advantage :

• These technique provides the accurate results
• Modifications are done to undergo the colony formation
• Accurately predicts the drug resistant tumors

Disadvantage :

• It has very low percent of dis aggregated tumor causing cells
• The process requires longer time i.e time consuming process
• It is very difficult to add the drugs sequentially during the process of testing
• It has another problem with the quality control