After blocking, a dilute solution of primary antibody (generally between 0.5 and 5 micrograms/mL) is incubated with the membrane under gentle agitation.
A product reminiscent of yogurt would have been produced, which, through gentle agitation and the separation of curds from whey would have resulted in the production of cheese; the cheese being essentially a concentration of the major milk protein, casein, and milk fat.
The mix was incubated for two hours in the cold room with gentle agitation, the beads were pelleted by spinning briefly on a bench top Eppendorf centrifuge, washed four times with PBB containing no BSA, and the pellet was dried under vacuum for twenty minutes.
Each mixture was incubated at 4 C for one hour with gentle agitation.
Using the anti-capillary forceps, hold the dried grid face-down on the surface of the ethanolic PTA for 1 min with gentle agitation, drain on the surface, transfer to the surface of 95% ethanol to rinse for 15-30 sec, drain on filter paper and allow to air-dry.
The beads were re-suspended with 50 μl of the same buffer, and added into the fragment-probe mix and incubated at room temperature for 30 minutes with constant gentle agitation.
The muscularis mucosae is composed of several thin layers of smooth muscle fibers oriented in different ways which keep the mucosal surface and underlying glands in a constant state of gentle agitation to expel contents of glandular crypts and enhance contact between epithelium and the contents of the lumen.
The slide was incubated in a static incubator at 42 C for 12-16 h, and washed by placing in a 250 ml solution of 2X SSC and 0.1% SDS at 42 C for 5 minutes with gentle agitation provided by rotation of a magnetic stir bar.
The slide was transferred quickly to a solution of 0.1X SSC, and 0.1% SDS, incubated for 10 minutes at room temperature with gentle agitation, and washed 5 additional times in 0.1X SSC for 1 min.
To generate cells that constitutively expressed human IL-1β, skin was first harvested from a euthanized Wistar rat (Charles River Laboratories, Wilmington, MA, USA), minced with a scalpel, and digested for 2 hours at 37 C under gentle agitation with 0.2% clostridial collagenase.