Are the tissue factor (TF) and phospholipid (PL) compositions available?

Exact TF concentration is not available. The manufacturing process for TF/PLs is proprietary. A mixture of PL and TF is used, which form micelles of varying PL/TF concentrations and thrombin generation is then tested for release (not concentrations). Generally, TF is in the low pM range, and PLs consist of a mixture of phosphatidylcholine and phosphatidylserine.

Can I prepare my own controls?

Yes. However, Technoclone TGA controls are preselected for TGA use; care is taken to not induce microparticle formation. Using one’s own controls or another source might have differences in microparticles between plasmas.

Can thrombin generation of microparticles be determined?

Yes. This again will vary depending on the experimental goals as well as the source of microparticles. Trial and error may be needed in this situation. For example, cell-derived microparticles are not the same as plasma-derived microparticles. To measure thrombin generation from microparticles, isolation of the microparticles is required first, which can be completed by research protocols in the literature or using a Microparticle Filtration Unit such as the Ceveron® MFU 500.

Can Technothrombin® TGA measure thrombin generation of cells?

Yes. However, the experiment protocol will vary depending on the experimental goals, as thrombin generation depends on any trigger that initiates thrombin generation. In some cases, cells are used as the trigger in plasma and tested for their capacity to generate thrombin.

Can Technothrombin® TGA be used with other species samples besides human?

Yes. However, researchers develop their own protocols and often vary the initial plasma sample dilution. For example, the dilution factor may vary depending on the species, but some researchers have had success diluting mouse plasma 1:2 in TGA buffer and dog plasma 1:3. This requires validation in the researcher’s laboratory.

If using a BioTek® fluorescent reader, can I keep the machine on all day?

Unfortunately, no, Technoclone and BioTek® recommend running only 2 consecutive runs and to switch the reader off for ~2-3 hours before starting the next run. The intensity of the lamp begins to diminish after too long. In addition, the bulb should be changed once a year (lifespan is ~500 hours anytime the instrument is on).

I received a BioTek® “Run-time error ‘1004’ Cannot insert object.” Now what?

This can occur with the Gen5 software associated with the BioTek® reader. Some newer versions of Microsoft Excel may be incompatible with older versions of Gen5 Software. Gen5 v1.11 or higher is needed. There is a cost associated with an upgrade to version 2.0, and BioTek® Customer Service must be contacted for this software upgrade. However, Gen5 v1.11 will work and can be downloaded here:
(Your Gen5 Software serial number will be needed).

I received another BioTek® error when trying to power export. What should I do?

While some issues may require your IT support, in some situations simply saving the data, and then restarting the computer, opening the data, and selecting power export again will fix the error (eg, this may resolve errors related to ‘cannot find path’). In addition, be sure that screen savers are turned off as this can interrupt the experiment.

I receive an error after BioTek® power export, what happened?

It’s likely that the data was not correctly copied into the excel evaluation template (Note: even provides document instructions on this). Cells must be copied to corresponding cell for data analysis of the calibration curve or samples/controls within the ‘Raw Data’ excel sheet tab. Raw data can then be analyzed with Excel protected formulas.

The BioTek® instrument reading is completed, how do I analyze my data?

You need to select ‘Power Export’. This will automatically open the data in an Excel file. However, as noted above, do not analyze data using standard Microsoft Excel. Your data must be copied and pasted into the Excel Evaluation Template/Software that corresponds to the instrument used.

I have created the protocols for the calibration curve and samples/controls, now what?

Prepare for running the experiment by opening the fluorescent reader software, creating a new experiment, and selecting the appropriate protocol. For some instruments, pre-heating should begin right away. To run the actual experiment, follow the short instructions provided with the Technothrombin® TGA for the calibration curve and samples/protocol. At some point, the plate layout within the Excel Evaluation Software/Template should be completed in order to help with subsequent data analysis.
Note: Only one calibration curve has to be done for each lot of substrate.

How do I set-up the protocol on the BioTek® instrument?

Protocol set-up on the instrument software only needs to be completed once. Two protocols are required: one for the calibration curve and one for running the samples and controls. Instrument read settings are critical! Technoclone maintains instrument application documents for the available instruments. The instrument application documents contain screenshots to help with set-up but the overall basic parameters settings will remain similar if the instrument software is ever updated.

Where is the TGA evaluation software?

The manufacturer refers to the Microsoft Excel data analysis templates as evaluation software. These are templates created by Technoclone that can be used for data analysis with a number of fluoresecent readers. The excel templates are available at or you can request one at

Do not analyze your data with standard Microsoft Excel.

What type of Fluorescent Reader do I need?

While the manufacturer currently recommends using the Ceveron® alpha TGA analyzer, alternative fluorescent readers can be used. Excel data evaluation templates have been created for the BioTek® FLx800™, Spectramax M5, BMG FLUOstar, Perkin Elmer Wallac Viktor, TECAN, and Thermo Fluoroskan.

Protein concentrations in plasma


Weight kDa



Fibrinogen 330 3000 9
Prothrombin 72 150 2
Factor V 330 20 0.05
Factor VII 50 0.5 0.01
Factor VIII 330 0.1 0.0003
Factor IX 56 5 0.09
Factor X 59 8 0.13
Factor XI 160 5 0.03
Factor XII 80 30 0.4
Factor XIII 320 10 0.03
Protein C 62 4 0.06
Protein S 70 10 (free) 0.14
Protein Z 62 2 0.03
Prekallikrein 86 50 0.6
HMW kininogen 120 70 0.6
Fibronectin 450 300 0.7
Plasminogen 92 200 2
t-PA 60 0.005 0.0001
Urokinase 53 0.004 0.0001
Antithrombin 58 145 2.5
Heparin Cofactor II 66 80 1.2
Plasmin Inhibitor 63 60 1
Protein C Inhibitor 57 4 0.07
α2-Macroglobulin 725 2000 3