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- 18:52, 17 January 2025 Restriction Digest (hist | edit) [968 bytes] Akipp4 (talk | contribs) (Created page with " Protocol 1. Check the NEB (or other supplier) website to confirm enzyme compatibility 2. In a PCR tube add the reaction components according to the table listed below 3. Incubate at 37°C for 30 min if time-saver qualified, 60 minutes if not Example Reaction Component Volume ddH2O Up to 50 µL 10X rCutSmart Buffer 5 µL DNA x (1 µg) Enzyme 1 1 µL Enzyme 2 1 µL Notes - NEB High-Fidelity (HF) enzymes are ordered whenever possible, they all use rCutSmart reactio...")
- 19:02, 29 July 2024 THP1 Culture and Differentiation (hist | edit) [3,089 bytes] Admin (talk | contribs) (Created page with "==== Maintenance ==== THP1 cells ([https://www.atcc.org/products/tib-202?matchtype=&network=x&device=c&adposition=&keyword=&gad_source=1&gclid=Cj0KCQjw-5y1BhC-ARIsAAM_oKlZTKs5zplX22cd1pVZB7E0-Cq7fDL5pcruuijrK0PqV9a-5LG5l24aAoEqEALw_wcB ATCC TIB-202]) should be maintained in RPMI + 10% FBS, with cells split 1:5 into fresh medium when the cell density reaches 1 x 10<sup>6</sup> cells/mL. Do not allow cell density to exceed 1.5 x 10<sup>6</sup> cells/mL. '''Classical Mac...") Tag: Visual edit
- 15:12, 10 July 2024 Frustrated Phagocytosis (hist | edit) [1,756 bytes] Admin (talk | contribs) (Created page with "==== Before the Experiment ==== # Prepare substrates and place them into the wells of a 12-well plate. These can be prepared by coating slides with a ligand of interest, or by microprinting. # Transfect cells if required, as per usual protocols. # Split macrophages into wells without coverslips, 1 well/frustrated phagocytosis assay (~250,000 cells/well). ==== Experiment: Fixed-Cell ==== # Pre-warm the substrates and centrifuges to 37C. # I...") Tag: Visual edit
- 16:43, 19 June 2024 Micropatterning Proteins (hist | edit) [2,222 bytes] Admin (talk | contribs) (Created page with "This protocol is based on IgG micropatterning methods as published by Freeman ''et al''. <ref>Freeman et al., (2016) “Integrins Form an Expanding Diffusional Barrier That Coordinates Phagocytosis.” Cell. V164#1, pp 128-40. http://linkinghub.elsevier.com/retrieve/pii/S0092867415015676</ref>It may need to be optimized for other proteins. ==== Preparation of the Microprinting Stamp. ==== This is optimized for a PDMS based stamp and may have to be modified for stamps m...") Tag: Visual edit