https://zylalab.org/protocols/ Recent content in Protocols on Zyla Lab Hugo en-us Sun, 28 Dec 2025 00:00:00 +0000 https://zylalab.org/protocols/go-grids/ Sun, 28 Dec 2025 00:00:00 +0000 https://zylalab.org/protocols/go-grids/ <p>This protocol describes how to prepare <strong>1-3 layers of semi-amorphous graphene oxide (GO)</strong> on Quantifoil grids, suitable for cryo-EM. GO films demonstrate reduced inelastic scattering and lower background compared to amorphous carbon, which improves phase contrast for imaging smaller complexes. In addition, it allows working with lower protein concentration and avoid prefered orientation. This protocol ensures 40-80% coverage of GO after vitrification, with precise handling steps to minimize aggregation and variation in coverage.</p> https://zylalab.org/protocols/mab-digest/ Sun, 28 Dec 2025 00:00:00 +0000 https://zylalab.org/protocols/mab-digest/ <p>This protocol details the steps for digesting 1 mg of immunoglobulin G (IgG) antibody using papain. It ensures efficient generation of Fab and Fc fragments with precise reagent concentrations and reaction conditions.</p> <hr> <h2 id="materials"><strong>Materials</strong></h2> <ol> <li> <p><strong>Buffers and Reagents</strong></p> <ul> <li><strong>1M Sodium Acetate, pH 5.5</strong></li> <li><strong>IgG Antibody</strong> (≥1 mg/mL)</li> <li><strong>100 mM L-Cysteine</strong> (prepared fresh)</li> <li><strong>500 mM Iodoacetamide</strong> (prepared fresh)</li> <li><strong>Papain Enzyme</strong> (~25 mg/mL)</li> <li><strong>500 mM EDTA, pH ~7.5</strong></li> </ul> </li> <li> <p><strong>Other Reagents</strong></p> <ul> <li><strong>Glacial Acetic Acid</strong></li> <li><strong>Distilled Deionized Water (ddH₂O)</strong></li> </ul> </li> <li> <p><strong>Consumables</strong></p>