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PCNA AI-CADD molecule thermal shift assay screening hits . AI-CADD screening: Left , plot of normalized melt curves of compounds + PCNA melt. Center , normalized derivative RFU of melt curve data. The DMSO control is plotted in black dashed lines. Right , . Calculated Δ Tm values of 19 AI-CADD hits. Thermal shift assays were performed using a Bio-Rad <t>CFX</t> <t>Connect</t> <t>Real-Time</t> <t>PCR</t> Detection System. and the final concentration of recombinant purified PCNA was 9 μM, and the final concentration of each compound was 1 mM. The reaction plates were heated from 25 °C to 95 °C with heating increments of 0.5 °C/min. Fluorescence intensity was measured within the excitation/emission ranges 470–505/540–700 nm. A negative control of 100% DMSO was included in each assay to obtain an apo- PCNA melting temperature (Tm apo ). The melting temperature of each reaction well was extrapolated from the midpoint of excitation of the produced melt curves by the CFX Maestro Software (Bio-Rad). The thermal shift values were calculated using the following equation: ΔTm = Tm + compound – Tm apo .
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PCNA AI-CADD molecule thermal shift assay screening hits . AI-CADD screening: Left , plot of normalized melt curves of compounds + PCNA melt. Center , normalized derivative RFU of melt curve data. The DMSO control is plotted in black dashed lines. Right , . Calculated Δ Tm values of 19 AI-CADD hits. Thermal shift assays were performed using a Bio-Rad <t>CFX</t> <t>Connect</t> <t>Real-Time</t> <t>PCR</t> Detection System. and the final concentration of recombinant purified PCNA was 9 μM, and the final concentration of each compound was 1 mM. The reaction plates were heated from 25 °C to 95 °C with heating increments of 0.5 °C/min. Fluorescence intensity was measured within the excitation/emission ranges 470–505/540–700 nm. A negative control of 100% DMSO was included in each assay to obtain an apo- PCNA melting temperature (Tm apo ). The melting temperature of each reaction well was extrapolated from the midpoint of excitation of the produced melt curves by the CFX Maestro Software (Bio-Rad). The thermal shift values were calculated using the following equation: ΔTm = Tm + compound – Tm apo .
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PCNA AI-CADD molecule thermal shift assay screening hits . AI-CADD screening: Left , plot of normalized melt curves of compounds + PCNA melt. Center , normalized derivative RFU of melt curve data. The DMSO control is plotted in black dashed lines. Right , . Calculated Δ Tm values of 19 AI-CADD hits. Thermal shift assays were performed using a Bio-Rad <t>CFX</t> <t>Connect</t> <t>Real-Time</t> <t>PCR</t> Detection System. and the final concentration of recombinant purified PCNA was 9 μM, and the final concentration of each compound was 1 mM. The reaction plates were heated from 25 °C to 95 °C with heating increments of 0.5 °C/min. Fluorescence intensity was measured within the excitation/emission ranges 470–505/540–700 nm. A negative control of 100% DMSO was included in each assay to obtain an apo- PCNA melting temperature (Tm apo ). The melting temperature of each reaction well was extrapolated from the midpoint of excitation of the produced melt curves by the CFX Maestro Software (Bio-Rad). The thermal shift values were calculated using the following equation: ΔTm = Tm + compound – Tm apo .
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PCNA AI-CADD molecule thermal shift assay screening hits . AI-CADD screening: Left , plot of normalized melt curves of compounds + PCNA melt. Center , normalized derivative RFU of melt curve data. The DMSO control is plotted in black dashed lines. Right , . Calculated Δ Tm values of 19 AI-CADD hits. Thermal shift assays were performed using a Bio-Rad <t>CFX</t> <t>Connect</t> <t>Real-Time</t> <t>PCR</t> Detection System. and the final concentration of recombinant purified PCNA was 9 μM, and the final concentration of each compound was 1 mM. The reaction plates were heated from 25 °C to 95 °C with heating increments of 0.5 °C/min. Fluorescence intensity was measured within the excitation/emission ranges 470–505/540–700 nm. A negative control of 100% DMSO was included in each assay to obtain an apo- PCNA melting temperature (Tm apo ). The melting temperature of each reaction well was extrapolated from the midpoint of excitation of the produced melt curves by the CFX Maestro Software (Bio-Rad). The thermal shift values were calculated using the following equation: ΔTm = Tm + compound – Tm apo .
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PCNA AI-CADD molecule thermal shift assay screening hits . AI-CADD screening: Left , plot of normalized melt curves of compounds + PCNA melt. Center , normalized derivative RFU of melt curve data. The DMSO control is plotted in black dashed lines. Right , . Calculated Δ Tm values of 19 AI-CADD hits. Thermal shift assays were performed using a Bio-Rad <t>CFX</t> <t>Connect</t> <t>Real-Time</t> <t>PCR</t> Detection System. and the final concentration of recombinant purified PCNA was 9 μM, and the final concentration of each compound was 1 mM. The reaction plates were heated from 25 °C to 95 °C with heating increments of 0.5 °C/min. Fluorescence intensity was measured within the excitation/emission ranges 470–505/540–700 nm. A negative control of 100% DMSO was included in each assay to obtain an apo- PCNA melting temperature (Tm apo ). The melting temperature of each reaction well was extrapolated from the midpoint of excitation of the produced melt curves by the CFX Maestro Software (Bio-Rad). The thermal shift values were calculated using the following equation: ΔTm = Tm + compound – Tm apo .
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PCNA AI-CADD molecule thermal shift assay screening hits . AI-CADD screening: Left , plot of normalized melt curves of compounds + PCNA melt. Center , normalized derivative RFU of melt curve data. The DMSO control is plotted in black dashed lines. Right , . Calculated Δ Tm values of 19 AI-CADD hits. Thermal shift assays were performed using a Bio-Rad CFX Connect Real-Time PCR Detection System. and the final concentration of recombinant purified PCNA was 9 μM, and the final concentration of each compound was 1 mM. The reaction plates were heated from 25 °C to 95 °C with heating increments of 0.5 °C/min. Fluorescence intensity was measured within the excitation/emission ranges 470–505/540–700 nm. A negative control of 100% DMSO was included in each assay to obtain an apo- PCNA melting temperature (Tm apo ). The melting temperature of each reaction well was extrapolated from the midpoint of excitation of the produced melt curves by the CFX Maestro Software (Bio-Rad). The thermal shift values were calculated using the following equation: ΔTm = Tm + compound – Tm apo .

Journal: Current Research in Structural Biology

Article Title: Screening for novel chemical scaffolds targeting PCNA identifies the Hsp90alpha inhibitor SNX-2112

doi: 10.1016/j.crstbi.2026.100183

Figure Lengend Snippet: PCNA AI-CADD molecule thermal shift assay screening hits . AI-CADD screening: Left , plot of normalized melt curves of compounds + PCNA melt. Center , normalized derivative RFU of melt curve data. The DMSO control is plotted in black dashed lines. Right , . Calculated Δ Tm values of 19 AI-CADD hits. Thermal shift assays were performed using a Bio-Rad CFX Connect Real-Time PCR Detection System. and the final concentration of recombinant purified PCNA was 9 μM, and the final concentration of each compound was 1 mM. The reaction plates were heated from 25 °C to 95 °C with heating increments of 0.5 °C/min. Fluorescence intensity was measured within the excitation/emission ranges 470–505/540–700 nm. A negative control of 100% DMSO was included in each assay to obtain an apo- PCNA melting temperature (Tm apo ). The melting temperature of each reaction well was extrapolated from the midpoint of excitation of the produced melt curves by the CFX Maestro Software (Bio-Rad). The thermal shift values were calculated using the following equation: ΔTm = Tm + compound – Tm apo .

Article Snippet: Thermal shift assays were performed using a Bio-Rad CFX Connect Real-Time PCR Detection System. and the final concentration of recombinant purified PCNA was 9 μM, and the final concentration of each compound was 1 mM.

Techniques: Thermal Shift Assay, Control, Real-time Polymerase Chain Reaction, Concentration Assay, Recombinant, Purification, Fluorescence, Negative Control, Produced, Maestro Software

PCNA AI-CADD molecule thermal shift assay screening hits . AI-CADD screening: Left , plot of normalized melt curves of compounds + PCNA melt. Center , normalized derivative RFU of melt curve data. The DMSO control is plotted in black dashed lines. Right , . Calculated Δ Tm values of 19 AI-CADD hits. Thermal shift assays were performed using a Bio-Rad CFX Connect Real-Time PCR Detection System. and the final concentration of recombinant purified PCNA was 9 μM, and the final concentration of each compound was 1 mM. The reaction plates were heated from 25 °C to 95 °C with heating increments of 0.5 °C/min. Fluorescence intensity was measured within the excitation/emission ranges 470–505/540–700 nm. A negative control of 100% DMSO was included in each assay to obtain an apo- PCNA melting temperature (Tm apo ). The melting temperature of each reaction well was extrapolated from the midpoint of excitation of the produced melt curves by the CFX Maestro Software (Bio-Rad). The thermal shift values were calculated using the following equation: ΔTm = Tm + compound – Tm apo .

Journal: Current Research in Structural Biology

Article Title: Screening for novel chemical scaffolds targeting PCNA identifies the Hsp90alpha inhibitor SNX-2112

doi: 10.1016/j.crstbi.2026.100183

Figure Lengend Snippet: PCNA AI-CADD molecule thermal shift assay screening hits . AI-CADD screening: Left , plot of normalized melt curves of compounds + PCNA melt. Center , normalized derivative RFU of melt curve data. The DMSO control is plotted in black dashed lines. Right , . Calculated Δ Tm values of 19 AI-CADD hits. Thermal shift assays were performed using a Bio-Rad CFX Connect Real-Time PCR Detection System. and the final concentration of recombinant purified PCNA was 9 μM, and the final concentration of each compound was 1 mM. The reaction plates were heated from 25 °C to 95 °C with heating increments of 0.5 °C/min. Fluorescence intensity was measured within the excitation/emission ranges 470–505/540–700 nm. A negative control of 100% DMSO was included in each assay to obtain an apo- PCNA melting temperature (Tm apo ). The melting temperature of each reaction well was extrapolated from the midpoint of excitation of the produced melt curves by the CFX Maestro Software (Bio-Rad). The thermal shift values were calculated using the following equation: ΔTm = Tm + compound – Tm apo .

Article Snippet: Thermal shift assays were performed using a BioRad CFX Connect Real-Time PCR Detection System.

Techniques: Thermal Shift Assay, Control, Real-time Polymerase Chain Reaction, Concentration Assay, Recombinant, Purification, Fluorescence, Negative Control, Produced, Maestro Software