Cell proliferation, meaning an increase in cell number, is a dynamic process balanced by cell division and cell loss. For the development of immune checkpoint inhibitors, the accurate measurement of T cell proliferation allows for a better understanding of the dynamics of cellular responses to extracellular stimuli. Thus, T cell proliferation assay is an important part of immune checkpoint drug development. Having focused on immune checkpoint drug development for more than ten years, Creative Biolabs has accumulated extensive immune checkpoint assay experience and has developed a comprehensive technology platform to satisfy every customers' different requirements. Our mature technique system and professional expert team are full guarantees for the quality of our services.
MTT assay is based on the conversion of MTT ((3-[4,5- dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) to insoluble formazan crystals by mitochondrial NAD(p)H-dependent oxidoreductase enzymes released in living cells. Proliferating cells possess a higher rate of MTT converting reaction while dead or slow-growing cells have low metabolism resulting in lower levels of MTT reduction. MTT assay leads us to determine the number of viable cells by measuring mitochondrial activity related to the number of formazan crystals. A spectrophotometer can measure formazan concentrations between 540 to 720 nm.
Fig.1 MTT assay. (Adan, 2016)
Alamar blue, also known as resazurin, is a non-toxic cell-permeable vital dye. Upon entering the cell, it is reduced to resorufin, which is red and highly fluorescent. A higher reduction rate of resazurin to resorufin in proliferating cells and a lower rate of this reduction in dead cells could be a marker to distinguish cellular viability. The metabolic reactions generating fluorescence signals can be measured at 570 and 630 nm.
ATP assay is based on monitoring ATP levels via bioluminescent detection. Natural firefly luciferase and luciferin react with cellular ATP and cause the production of a certain level of light that can be measured by a luminometer. The amount of luminescent light is directly proportional to the viability of cells. ATP assay is advantageous compared to other methods with different ranges such as short time protocol, high sensitivity, and large usage areas. The signal is also present for a long time, which makes the measuring step easier.
Fig.2 The principle of ATP assay. (Adan, 2016)
Besides the three methods mentioned above, the technology platform at Creative Biolabs has other assays for choice to meet customers' different demands. Here we listed the most common T cell proliferation assays that are serving for reference in the below chart. If you are unsure which method is the most suitable for your research, please contact our expert team who can help you design the solution that suits your research best.
|Method||Principle of Action||Advantages|
|Trypan Blue||Membrane integrity||
Easy to apply
Proper for potential drug follow-up studies
|Alamar Blue||Cell metabolism||
Suitable for long-term studies
Highly stable in the culture medium
|LDH Release Assay||Cell membrane integrity & Cell metabolism||
Accepted absorbance or fluorescent options
|G6PD Release Assay||Cell membrane integrity & Cell metabolism||
More sensitive than LDH
More efficient than LDH
Short term protocol
|ATP Assay||ATP production||
Short term protocol
Very good sensitivity
Large usage areas
|Sulforhodamine B Assay||Cellular protein content||
Suitable for high-throughput screening
Proper for long-term studies
High sensitivity in low cell numbers
As a professional immune checkpoint assay service provider, Creative Biolabs has a comprehensive platform with advanced facilities. Thus, we are capable of providing high-quality T cell proliferation assay services to global clients. If you are interested in T cell proliferation assay services or any other services on our website, please feel free to contact us for more information.
All listed customized services & products are for research use only, not intended for pharmaceutical, diagnostic, therapeutic, or any in vivo human use.
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