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Sypro orange dye protein thermal shift dye

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SYPRO Orange dye is a protein thermal shift dye used to monitor protein unfolding during thermal denaturation experiments. It is a fluorescent dye that binds to hydrophobic regions of proteins and shows an increase in fluorescence upon binding. The dye can be used to study protein stability and ligand binding interactions.

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Lab products found in correlation

Transparent 384 well plates, by Thermo Fisher Scientific (2 mentions) Fxr lbd, by Thermo Fisher Scientific (2 mentions) Pts software, by Thermo Fisher Scientific (2 mentions) Quantstudio 6 flex real time qpcr system, by Thermo Fisher Scientific (2 mentions)

Spelling variants of this product

SYPRO Orange (691 citations) SYPRO Orange dye (340 citations) Protein Thermal Shift Dye (61 citations) SYPRO Orange Protein Thermal Shift Dye (3 citations) Thermal shift dye (3 citations)

2 protocols using sypro orange dye protein thermal shift dye

1

Thermal Shift Assay for FXR-LBD

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
IsoDCA and CDCA were tested for their ability to alter the melting temperature (Tm) of recombinant FXR-LBD (Invitrogen) using a modified version of the protocol established by Schmidt et al.27 (link) Briefly, BAs and FXR-LBD (final concentration: 500 nM) were diluted in assay buffer [10 mM TRIS (pH 8.3), 0.5 mM EDTA, 100 mM NaCl and 5 mM DTT, added fresh before assay from a 100x frozen stock] and combined into transparent 384 well plates (Applied Biosystems). SYPRO Orange dye (Protein Thermal Shift Dye, Applied Biosystems) was added at a final concentration of 1: 2,000 in a reaction volume of 20 μL. Temperature-dependent changes in fluorescence were detected in a QuantStudio 6 Flex Real-time qPCR System (Applied Biosystems). The equipment was programmed with the following thermal profile: Step 1, Temp: 25 °C, Time: 2 minutes/ Step 2, Temp: 99 °C, Time: 2 minutes /Ramp mode: Continuous /Ramp rate: Step 1: 1.6°C /s, Step 2: 0.05°C /s. Data analysis was performed using the PTS software (Applied Biosystems).
Campbell C., McKenney P.T., Konstantinovsky D., Isaeva O.I., Schizas M., Verter J., Mai C., Jin W.B., Guo C.J., Violante S., Ramos R.J., Cross J.R., Kadaveru K., Hambor J, & Rudensky A.Y. (2020). Bacterial metabolism of bile acids promotes peripheral Treg cell generation. Nature, 581(7809), 475-479.
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2

Thermal Shift Assay for FXR-LBD

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
IsoDCA and CDCA were tested for their ability to alter the melting temperature (Tm) of recombinant FXR-LBD (Invitrogen) using a modified version of the protocol established by Schmidt et al.27 (link) Briefly, BAs and FXR-LBD (final concentration: 500 nM) were diluted in assay buffer [10 mM TRIS (pH 8.3), 0.5 mM EDTA, 100 mM NaCl and 5 mM DTT, added fresh before assay from a 100x frozen stock] and combined into transparent 384 well plates (Applied Biosystems). SYPRO Orange dye (Protein Thermal Shift Dye, Applied Biosystems) was added at a final concentration of 1: 2,000 in a reaction volume of 20 μL. Temperature-dependent changes in fluorescence were detected in a QuantStudio 6 Flex Real-time qPCR System (Applied Biosystems). The equipment was programmed with the following thermal profile: Step 1, Temp: 25 °C, Time: 2 minutes/ Step 2, Temp: 99 °C, Time: 2 minutes /Ramp mode: Continuous /Ramp rate: Step 1: 1.6°C /s, Step 2: 0.05°C /s. Data analysis was performed using the PTS software (Applied Biosystems).
Campbell C., McKenney P.T., Konstantinovsky D., Isaeva O.I., Schizas M., Verter J., Mai C., Jin W.B., Guo C.J., Violante S., Ramos R.J., Cross J.R., Kadaveru K., Hambor J, & Rudensky A.Y. (2020). Bacterial metabolism of bile acids promotes peripheral Treg cell generation. Nature, 581(7809), 475-479.
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