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dsf buffer (Thermo Fisher)

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Thermo Fisher dsf buffer
Dsf Buffer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/dsf+buffer/Phosphate/pmc12657311-40-8-31
Average 99 stars, based on 1 article reviews

dsf buffer - by Bioz Stars, 2026-09

99/100 stars

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Purification:

Article Title: Allosteric inhibition rescues hydrocephalus caused by catalytically inactive Shp2
Article Snippet: The thermostability of each protein was measured using differential scanning fluorimetry (DSF) on MicroAmp Fast Optical 96-well Reaction plates (Applied Biosystems, 4346906) at working volumes of 20 μL. .. Purified proteins were diluted in DSF buffer (20 mM HEPES pH 7.5, 50 mM NaCl, 0.4% DMSO) to 10 μM in a mixture that also contained SYPRO Orange Protein Gel Stain (Thermo Fisher, S-6650), diluted 200-fold from a 5000x stock to 25x as the working concentration. .. Melting curves were measured on an Applied Biosystems Step-One Plus RT-PCR thermocycler between 25 °C and 95 °C with a gradient of +0.5 °C per minute (excitation: 472 nm; emission: 570 nm).

Staining:

Article Title: PK11007 Covalently Inhibits Thioredoxin Reductase 1 to Induce Oxidative Stress and Autophagy Impairment in NSCLC Cells
Article Snippet: .. 3 μM TXNRD1 was incubated with 100 μM NADPH and 100 μM compounds, including PK11007, APR-246, MQ and TRi-1 in a DSF buffer (20 mM HEPES, 100 mM NaCl, pH 7.4) at room temperature for 1 h. Subsequently, SYPRO Orange stain (S6650, Thermo Fisher, Waltham, MA, USA) was added to achieve a final concentration of 5×. ..

Concentration Assay:

Article Title: CdmR, a MarR family protein from Clostridioides difficile , binds with positive cooperativity in the predicted promoter region of a MATE efflux transporter
Article Snippet: Thermal denaturation studies were carried out in a StepOne Real-Time PCR System equipped with a 48-well block (Applied Biosystems, USA). .. Each sample contained 3 μ g protein in DSF buffer (50 mM sodium phosphate (pH 7.5), 50 mM NaCl, 0.2 mM BME, and a 1x concentration of Protein Thermal Shift Dye (Applied Biosystems, USA)) at a final volume of 20 μ L. Data were fit to a Boltzmann function assuming a two-state protein unfolding transition according to the procedure described by Ref. [ ] to determine protein T m , Δ G u ° , Δ H u ° , and Δ S u ° . ..

Article Title: Phenomics‐Based Discovery of Novel Orthosteric Choline Kinase Inhibitors
Article Snippet: .. DSF buffer (20 mM HEPES, 150 mM NaCl, 0.5 mM TCEP, pH 7.5) containing 5x SYPRO Orange (Thermo Fisher Scientific) and CHKA (2 μM) was incubated with 10 μM compound at 2% DMSO final concentration in a 96-well semi-skirted real time PCR plate. .. Fluorescence was measured in each well over a 40-80 °C gradient using a Stratagen MX3005p qPCR instrument (Agilent Technologies).

other:

Article Title: Deep mutational scanning of the multi-domain phosphatase SHP2 reveals mechanisms of regulation and pathogenicity.
Article Snippet: The cleavagemixture was applied through2mLofNi-NTAgravity column (ThermoFisher) to remove uncleaved protein andTEV protease, and the flow throughwas concentrated to less than 1mL.

Incubation:

Real-time Polymerase Chain Reaction:

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$(a) UNC10088 C -induced conformational change of the loop adjacent to C326. Structural overlay of the SCF FBXO22 -UNC10088 C -NSD2 PWWP1 model (dark gray) and the SCF FBXO22 -BACH1 BTB complex (light gray, PDB: 8UA3) , revealing the displacement of FBXO22 residues Y390. (b) Chemical structures of UNC8732 analogs that contain an aromatic aldehyde in place of the aliphatic amine of UNC8732. (c) Benzaldehyde derivatives UNC10415667 and UNC10415668 promote FBXO22-dependent ubiquitination of *NSD2 PWWP1 when added at 0.5 µM, based on fluorescent scanning of an SDS-PAGE gel. (d) Treatment of U2OS NSD2-HiBit cells with UNC10415667 (1 µM) for 6 hours resulted in a potent reduction of NSD2 levels by about 75%. A more modest effect was observed upon treatment with UNC10415668, and all other compounds had no significant effect. n ≥ 3 independent experiments. Error bars: standard error of the mean. (e) Treatment of U2OS NSD2 HiBit cells with UNC10415667 in a dose response fashion revealed a DC 50 value of 0.46 µM, which is about 2-fold more potent than UNC8732 and UNC10088. n = 3 independent experiments. Error bars: standard error of the mean. (f) UNC10415667 uses C326 to recruit NSD2 to the FBXO22 FIST domain as monitored by size-exclusion chromatography. Chromatograms of different elution profiles between mixtures of WT FBXO22 FIST or a C326A mutant with NSD2 and UNC10415667 (top). Representative Coomassie-stained SDS-PAGE gels of collected fractions. (g) Differential scanning fluorimetry (DSF) was used to monitor the stability of FBXO22 FIST wild type and variants in the presence of UNC10088 or UNC10415667 compared to DMSO, revealing that both compounds stabilize WT FBXO22 FIST and the C326 single cysteine variant (C326 only) as evidenced by an increase in melting temperature (ΔT m ), but not the C326A mutant. n = 3 independent experiments. Error bars: standard error of the mean.$

Journal: bioRxiv

Article Title: Structural basis of NSD2 degradation via targeted recruitment of SCF-FBXO22

doi: 10.1101/2025.08.29.673087

Figure Lengend Snippet: (a) UNC10088 C -induced conformational change of the loop adjacent to C326. Structural overlay of the SCF FBXO22 -UNC10088 C -NSD2 PWWP1 model (dark gray) and the SCF FBXO22 -BACH1 BTB complex (light gray, PDB: 8UA3) , revealing the displacement of FBXO22 residues Y390. (b) Chemical structures of UNC8732 analogs that contain an aromatic aldehyde in place of the aliphatic amine of UNC8732. (c) Benzaldehyde derivatives UNC10415667 and UNC10415668 promote FBXO22-dependent ubiquitination of *NSD2 PWWP1 when added at 0.5 µM, based on fluorescent scanning of an SDS-PAGE gel. (d) Treatment of U2OS NSD2-HiBit cells with UNC10415667 (1 µM) for 6 hours resulted in a potent reduction of NSD2 levels by about 75%. A more modest effect was observed upon treatment with UNC10415668, and all other compounds had no significant effect. n ≥ 3 independent experiments. Error bars: standard error of the mean. (e) Treatment of U2OS NSD2 HiBit cells with UNC10415667 in a dose response fashion revealed a DC 50 value of 0.46 µM, which is about 2-fold more potent than UNC8732 and UNC10088. n = 3 independent experiments. Error bars: standard error of the mean. (f) UNC10415667 uses C326 to recruit NSD2 to the FBXO22 FIST domain as monitored by size-exclusion chromatography. Chromatograms of different elution profiles between mixtures of WT FBXO22 FIST or a C326A mutant with NSD2 and UNC10415667 (top). Representative Coomassie-stained SDS-PAGE gels of collected fractions. (g) Differential scanning fluorimetry (DSF) was used to monitor the stability of FBXO22 FIST wild type and variants in the presence of UNC10088 or UNC10415667 compared to DMSO, revealing that both compounds stabilize WT FBXO22 FIST and the C326 single cysteine variant (C326 only) as evidenced by an increase in melting temperature (ΔT m ), but not the C326A mutant. n = 3 independent experiments. Error bars: standard error of the mean.

Article Snippet: 9.5 μL of the master mix containing FBXO22 FIST WT or variants, SYPRO Orange Dye (Invitrogen), and DSF buffer (20 mM Tris 7.5, 150 mM NaCl, and 1 mM TCEP) were added to a 384-well qPCR plate (Genesee Scientific).

Techniques: Ubiquitin Proteomics, SDS Page, Size-exclusion Chromatography, Mutagenesis, Staining, Variant Assay