{"batchcomplete":"","continue":{"lecontinue":"20220302180138|148","continue":"-||"},"query":{"logevents":[{"logid":158,"ns":0,"title":"Restriction Digest","pageid":114,"logpage":114,"revid":252,"params":{},"type":"create","action":"create","user":"Akipp4","timestamp":"2025-01-17T18:52:51Z","comment":"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\u00b0C for 30 min if time-saver qualified, 60 minutes if not  Example Reaction\t  Component\tVolume ddH2O\tUp to 50 \u00b5L 10X rCutSmart Buffer\t5 \u00b5L DNA\tx (1 \u00b5g) Enzyme 1\t1 \u00b5L Enzyme 2\t1 \u00b5L  Notes -\tNEB High-Fidelity (HF) enzymes are ordered whenever possible, they all use rCutSmart reactio...\""},{"logid":157,"ns":0,"title":"THP1 Culture and Differentiation","pageid":113,"logpage":113,"revid":232,"params":{},"type":"create","action":"create","user":"Admin","timestamp":"2024-07-29T19:02:53Z","comment":"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...\""},{"logid":156,"ns":0,"title":"Frustrated Phagocytosis","pageid":112,"logpage":112,"revid":227,"params":{},"type":"create","action":"create","user":"Admin","timestamp":"2024-07-10T15:12:48Z","comment":"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 [[Micropatterning Proteins|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...\""},{"logid":155,"ns":0,"title":"Micropatterning Proteins","pageid":111,"logpage":111,"revid":223,"params":{},"type":"create","action":"create","user":"Admin","timestamp":"2024-06-19T16:43:38Z","comment":"Created page with \"This protocol is based on IgG micropatterning methods as published by Freeman ''et al''. <ref>Freeman et al., (2016) \u201cIntegrins Form an Expanding Diffusional Barrier That Coordinates Phagocytosis.\u201d 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...\""},{"logid":154,"ns":6,"title":"File:MagnetTool.png","pageid":110,"logpage":110,"revid":208,"params":{},"type":"create","action":"create","user":"Admin","timestamp":"2022-03-03T15:36:19Z","comment":""},{"logid":153,"ns":6,"title":"File:MagnetTool.png","pageid":110,"logpage":110,"revid":208,"params":{"img_sha1":"cq873hb9u2t9k8ujrvlm43u9g5p125l","img_timestamp":"2022-03-03T15:36:19Z"},"type":"upload","action":"upload","user":"Admin","timestamp":"2022-03-03T15:36:19Z","comment":""},{"logid":152,"ns":6,"title":"File:50mm BasePlate.png","pageid":109,"logpage":109,"revid":207,"params":{},"type":"create","action":"create","user":"Admin","timestamp":"2022-03-03T15:35:54Z","comment":""},{"logid":151,"ns":6,"title":"File:50mm BasePlate.png","pageid":109,"logpage":109,"revid":207,"params":{"img_sha1":"oer9m2lrjmzv4cezx61urmgzun0bol3","img_timestamp":"2022-03-03T15:35:54Z"},"type":"upload","action":"upload","user":"Admin","timestamp":"2022-03-03T15:35:54Z","comment":""},{"logid":150,"ns":6,"title":"File:18mm BasePlate.png","pageid":108,"logpage":108,"revid":206,"params":{},"type":"create","action":"create","user":"Admin","timestamp":"2022-03-03T15:35:35Z","comment":""},{"logid":149,"ns":6,"title":"File:18mm BasePlate.png","pageid":108,"logpage":108,"revid":206,"params":{"img_sha1":"fd8f16jbws0134wr41u2oie5jsr1szj","img_timestamp":"2022-03-03T15:35:35Z"},"type":"upload","action":"upload","user":"Admin","timestamp":"2022-03-03T15:35:35Z","comment":""}]}}