Author: Drummond, Sheona P.; Hildyard, John; Firczuk, Helena; Reamtong, Onrapak; Li, Ning; Kannambath, Shichina; Claydon, Amy J.; Beynon, Robert J.; Eyers, Claire E.; McCarthy, John E. G.
Title: Diauxic shift-dependent relocalization of decapping activators Dhh1 and Pat1 to polysomal complexes Document date: 2011_6_28
ID: 1jdcdwxo_10
Snippet: Polysomal gradient analysis was performed as described previously (18) . In Figure 3C , cell cultures were incubated for 1 h at 30 C with 1 mg/ml puromycin dihydrochloride (Melford) in the presence of a low concentration of lyticase to ensure maximal access to the translation machinery. Cells were then processed as above, or clarified lysate was treated with 1 mg/ml RNase A prior to loading onto sucrose gradients. To generate samples for Western .....
Document: Polysomal gradient analysis was performed as described previously (18) . In Figure 3C , cell cultures were incubated for 1 h at 30 C with 1 mg/ml puromycin dihydrochloride (Melford) in the presence of a low concentration of lyticase to ensure maximal access to the translation machinery. Cells were then processed as above, or clarified lysate was treated with 1 mg/ml RNase A prior to loading onto sucrose gradients. To generate samples for Western blotting, proteins were recovered from gradient fractions by TCA precipitation. TCM-tagged proteins in polysomal gradient fractions were labelled using a Lumio-Green Detection Kit (Invitrogen) according to manufacturer's instructions and visualized in-gel using a UV transilluminator. Bands on western blots were visualized using chemiluminescence. This study PTC327 MAT PAT1:TCM:kanMX ade2-1 ura3-1 leu2-3,112 his3-11,15 can1-100
Search related documents:
Co phrase search for related documents- cell culture and low concentration: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10
- cell culture and previously describe: 1
- cell culture and sucrose gradient: 1, 2, 3
- cell culture and translation machinery: 1, 2, 3, 4
- cell culture and western blot: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42
- cell culture and western blotting: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26
- cell culture and western blotting sample: 1
- cell process and Detection kit: 1
- cell process and low concentration: 1
- cell process and western blot: 1, 2, 3, 4, 5, 6
- cell process and western blotting: 1, 2, 3
- Detection kit and Green Detection kit: 1
- Detection kit and low concentration: 1, 2, 3, 4, 5
Co phrase search for related documents, hyperlinks ordered by date