low profile plates  (Hampton Research Corp)

Journal: Biotechnology for Biofuels and Bioproducts

Article Title: There is an “I” in team: individual improvements in supercharged cellulase cocktail facilitates cooperative cellulose degradation

doi: 10.1186/s13068-026-02740-y

Figure Lengend Snippet: Thermodynamic decoupling of CBM and catalytic-domain melting in D5 CBM2a–WT Cel6B retains catalytic-domain thermostability. Enzyme melt curves comparing the native enzyme (dashed black line) to purified ( A ) D3 CBM2a–WT Cel6B, ( B ) D5 CBM2a–WT Cel6B, ( C ) D7 CBM2a–WT Cel6B, and ( D ) D7 CBM2a–D5 Cel6B. Thermal shift assays were conducted using 5 µL of 50 × SYPRO orange dye, 2.5 µL of 1 M sodium phosphate pH 6.0, 17.5 µL of DI water, and 25 µL of 10 µM protein dilution (in DI water) in an optically clear 96-well PCR plate. Heat denaturation and subsequent fluorescent signal monitoring were done in a Bio-Rad CFX Opus 96 real-time PCR system by equilibrating samples first to 25 °C and heating to 99 °C with a 0.5 °C increment and 5 s read time. All proteins were tested in quadruplicates and data were processed in MATLAB to average replicates, subtract background fluorescence of a water blank, and smooth curves

Article Snippet: Briefly, 5 μL of 50 × SYPRO orange dye (Thermo Fischer Scientific), 2.5 μL of 1 M sodium phosphate buffer pH 6.0, 17.5 μL of DI water, and 25 μL of 10 μM protein dilution were added to a clear low-profile Hard-Shell 96-well PCR plate (Bio-Rad) and sealed with an optically clear adhesive Microseal “B” plate seal (Bio-Rad).

Techniques: Purification, Real-time Polymerase Chain Reaction, Fluorescence