Protein Thermal Shift™ Starter Kit
Protein Thermal Shift™ Starter Kit

Protein Thermal Shift™ Starter Kit

Applied Biosystems™ Protein Thermal Shift™ kits allow users to employ a dye that binds to exposed hydrophobic residues to monitorRead more
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Catalog NumberQuantity
4462263
Catalog number 4462263
Price (USD)
590.00
Each
Add to cart
Price (USD)
590.00
Each
Add to cart
Applied Biosystems™ Protein Thermal Shift™ kits allow users to employ a dye that binds to exposed hydrophobic residues to monitor the thermal stability of proteins using a real-time PCR instrument in order to identify buffer conditions that stabilize the protein of interest or screen libraries of ligands for compounds that bind the protein of interest.

Save Time and Money with Your Sample Screening
This solution significantly reduces the time and cost associated with screening samples relative to currently available methods. Protein Thermal Shift™ products that are run on our real-time PCR systems enable researchers to inexpensively and efficiently:

• Screen samples for relative protein thermal stability
• Screen molecules (e.g. small molecule drug candidates, fragment-based libraries, and antibodies) for ligand binding to proteins in a high-throughput manner
• Screen samples for stability changes after protein point mutations or modifications
• Systematically identify suitable⁄optimal buffer conditions for the measurement of protein-ligand interactions, production of stable protein preparations, protein crystallization, and storage of proteins
• Quickly QC protein preps to ensure protein integrity and⁄or purity

Two Kits to Choose From
The Protein Thermal Shift™ Dye Kit contains the dye and buffer for performing protein thermal shift experiments, enough for up to 1000 reactions. The Protein Thermal Shift™ Starter Kit contains the Dye Kit plus a control protein and control ligand for test experiments and troubleshooting. The Protein Thermal Shift™ Starter Kit components are packaged in two boxes, one to be stored at -20°C and the other at 4°C, for maximum flexibility in reagent formulation.

Simplify Analysis with Protein Thermal Shift™ Software
Our complete Protein Thermal Shift™ solution includes analysis software which is compatible with run files generated from five of our real-time PCR systems: StepOne™ Real-Time PCR System, StepOnePlus™ Real-Time PCR system, 7500 Fast Real-Time PCR System, and ViiA™7 Real-Time PCR System.
For Research Use Only. Not for use in diagnostic procedures.
Specifications
Detection MethodPrimer-probe
PCR MethodqPCR
TechniqueFluorescence Intensity
For Use With (Equipment)7500 Fast System, 7500 System, StepOne™, StepOnePlus™, ViiA™ 7 System
Kit TypeStarter Kit
Product LineProtein Thermal Shift™
Target SpecificityNot Target-Specific
For Use With (Application)Protein Thermal Shift
No. of Reactions1,000 reactions
Sample TypeProtein
Unit SizeEach
Contents & Storage
Store dye and buffer at 18° to 25°C
Store control ligand and control protein at -15° to -25°C.

Frequently asked questions (FAQs)

In the Protein Thermal Shift Assay, my replicates have different levels of fluorescence. Is this a problem?

We recommend looking at the spread in Tm, which is more important than the relative fluorescence.

What can I do for a protein that starts out high and then shows a region of melt (flattening of the curve, but no real rise in signal) when performing a Protein Thermal Shift assay?

Some proteins have hydrophobic residues on the surface and the dye binds to these residues. Heating results in unfolding of the protein causing more hydrophobic residues to be exposed. The dyes bind preferentially to these inner locations and so there is a flattening (or a very low rise) of the melt discernable in the melt profile. If there is no positive slope, you will not get a Boltzmann Tm, but you should still get a derivative one. And you can always draw a manual region to get a Tm out. Some proteins will not work with this technology if the hydrophobic residues are already exposed on the surface and the dye binds strongly to it. Please contact Technical Support at techsupport@thermofisher.com about the possibility of other dyes being available for this issue.

I am getting an error message when I try to open my *.eds data file in the Protein Thermal Shift Software? What should I do?

Make sure that you first open the file in the corresponding instrument software, click “Analyze”, and then save the file, before trying to open with the Protein Thermal Shift Software. The file must first be analyzed before it can be used in the Protein Thermal Shift Software.

The software allows for data from different plates to be analyzed together, what should be considered when mixing data from multiple plates?

The software will allow for ≥100 plates per study. We allow the user this flexibility but do not recommend you mix data from multiple plates unless they have validated their results in advance. At a minimum, we recommend researchers include a reference assay in each plate to ensure reproducibility.

Which analysis method should I choose? The Boltzmann fit or the derivative method?

We provide two independent methods because they each have unique things to offer in terms of the analysis. The two-state Boltzmann model has a physical meaning and appeal. It also provides a great way to normalize across noisy undulations in the signal. However, those undulations may be of actual interest and not noise, such as for multi-domain proteins where they may correspond to different domains coming apart in stages. Here the two-state model is inappropriate. The derivative method can help get a temperature at which the local peaks occur. These are two completely unrelated approaches. If the two-state model is a great fit for your data, the results should be in close agreement.

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