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General

What is the difference between Relative and Absolute Quantitation? Which type of quantitation is used for proteomic samples?

Relative quantitation compares the amount of an analyte in different samples. Absolute quantitation measures the actual amount of an analyte in a sample using a known standard at different concentrations. Label-free, SILAC, and TMT (Tandem Mass Tag) workflows are used for relative quantitation of protein samples. Heavy (i.e., AQUA) peptides are used as standard for absolute quantitation of target proteins.

What is the main purpose of Tandem Mass Tag (TMT) reagents?

Tandem Mass Tag technology is used to individually label different protein samples so they can be combined into a single sample for LC-MS analysis. The major advantages of this workflow are higher sample throughput, less instrument analysis time, high precision of peptide quantitation among replicates, and fewer missing quantified proteins among different samples.

What type of mass spectrometer can I use for Tandem Mass Tag (TMT) analysis?

We recommend using high-resolution Orbitrap Tribrid (e.g. Orbitrap Ascend or Orbitrap Eclipse), Orbitrap Exploris (e.g., 240, 480), or Q Exactive Plus series of instruments.

How many samples can I combine for simultaneous, multiplex proteomic mass spec quantitation?

SILAC can be used to multiplex 2 to 3 samples. TMT reagents can be used to multiplex 6 to 11 samples. TMTpro reagents can be used to measure up to 16 samples simultaneously.

What sample types can I use with different mass spec quantitation systems?

Label-free, TMT reagents, and TMTpro reagents can be used with any protein sample type. SILAC and Neucode quantitation can only be used with cell lines and model organisms that can be metabolically labeled.

Can I use TMT reagents in combination with TMTpro reagents?

TMT reagents and TMTpro reagents are different chemicals with different masses. Therefore, we do not recommend mixing the two tags as the combined samples would be significantly more complex resulting in fewer quantified proteins.

For Research Use Only. Not for use in diagnostic procedures.

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