目次
| サンプル種 | サンプル量 | 標的タンパク質 | ホルムアルデヒド処理 | UCD-250使用条件 | DNA長 | ChIP assay protocol |
|---|---|---|---|---|---|---|
| 1.5mL チューブ | ||||||
| プラスミド DNA を磁気ビーズに固定し、クロマチン作製 | plasmidDNA 400ng |
複製因子 | 5 min at RT in formaldehyde 1% final |
250 W, 30 sec ON/30 sec OFF/15 cycle (15 min) |
500-300 bp ladder | Upstate 社 protocol |
| Human B cell | 200 μL, 3 x106 cells |
アセチル化ヒストン | 5 min at RT in formaldehyde 1% final; |
250 W, 30 sec ON/1 min OFF/11cycle (16.5 min) |
2 kbp 以下 ladder | Upstate 社 protocol |
| 15mL チューブ | ||||||
| Hela cells | 1.4 mL, 2x107 cells |
TBP, RNA polymerase II, Suv39H1, Paf1 etc. |
10 min at RT in formaldehyde 0.75% |
250 W, 30 sec ON/30 sec OFF/20 cycle (20 min) |
800-250 bp ladder | Abcam 社 protocol |
| U2OS cells | 1.4 mL, 2x107 cells |
TBP, RNA polymerase II, Suv39H1, Paf1 etc. |
10 min at RT in formaldehyde 0.75% |
250 W, 30 sec ON/30 sec OFF/7 cycle (7 min) |
800-250 bp ladder | Abcam 社 protocol |
| HL60 | 1.4 mL, 2x107 cells |
TBP, RNA polymerase II, Suv39H1, Paf1 etc. |
10 min at RT in formaldehyde 0.75% |
250 W, 30 sec ON/30 sec OFF/20 cycle (20 min) |
800-250 bp ladder | Abcam 社 protocol |
| MCF7 | 1.4 mL, 2x107 cells |
TBP, RNA polymerase II, Suv39H1, Paf1 etc. |
10 min at RT in formaldehyde 0.75% |
250 W, 30 sec ON/30 sec OFF/25 cycle (25 min) |
800-250 bp ladder | Abcam 社 protocol |
| 酵母 (超音波処理前にビーズで破砕) | OD=10, 4mL | 転写因子 | 30 min at RT in formaldehyde 4% final |
250 W, 30 sec ON/1 min OFF/5 cycle (7.5 min) |
1000-500 bp ladder | |