Table 2.
Details of primary and secondary antibodies used during western blotting
| Antibodies | MW (Kd) |
Antibody dilutions |
||
|---|---|---|---|---|
| Primary | Secondary | Ref (primaries) | ||
| UBE1 (N-20) |
110 |
1:200 (Goat) |
1:50,000 (Sigma) |
Santa Cruz 47555 |
| E217Kb/UBE2D2 |
17 |
1:1,000 (Rb) |
1:50,000 (Sigma) |
S.S.Wing (UBC4) (gift) |
| LASU1 /URE-B1/E3 |
485 |
1:1,000 (Rb) |
1:50,000 (Sigma) |
S.S.Wing (HUWE1) (gift) |
| CDYL (F-15) |
66 |
1:50 (Goat) |
1:20,000 (Sigma) |
Santa Cruz 34147 |
| BRDT/brd6 |
75 |
1:100 (Goat) |
1:5,000 (s.c) |
Everest EB05796 |
| HDAC1(C-19) |
52 |
1:100 (Goat) |
1:50,000 (Sigma) |
Santa Cruz 6298 |
| TOPO IIβ (H-286) |
180 |
1:200 (Rb) |
1:5,000 (s.c) |
Santa Cruz 13059 |
| PEM (M-15) |
21-24 |
1:50 (Goat) |
1:4,000 (s.c) |
Santa Cruz 21650 |
| Proteasomes purified fish |
24-31 |
1:3,000 (Ms Mab) |
1:50,000 (Sigma) |
T. Tokumoto (GC3α) (gift) |
| 20S Proteasomeα1(C7) |
24-31 |
1:200 (Ms Mab) |
1:5,000 (s.c) |
Santa Cruz 166073 |
| 20S Proteasomeα5(H-53) |
24-31 |
1:250 (Rb) |
1:5,000 (s.c) |
Santa Cruz 67342 |
| 20S Proteasomeα7(MCP72) |
24-31 |
1:500 (Ms MAb) |
1:50,000 (Sigma) |
Santa Cruz 58417 |
| H1 |
17 |
1:20 (Ms Mab) |
1:4,000 (s.c) |
Abcam 62884 |
| H2A somatic |
14 |
1:20 (Rb) |
1:20,000 (Sigma) |
Biovision 3621-100 |
| H2Bsomatic |
14 |
1:3,000 (Rb) |
1:4,000 (s.c) |
Epitomics 1847-1 |
| H3 somatic |
15 |
1:1,000 (Rb) |
1:50,000 (Sigma) |
Santa Cruz 8654R |
| tH2B |
15 |
1:250 (Rb) |
1:25,000 (Sigma) |
Millipore 07680 |
| H4 |
11 |
1:20 (Rb) |
1:20,000 (Sigma) |
Biovision 3624-100 |
| H2AUb |
25 |
1:65 (Ms Mab/IgM) |
1:8,500 (anti-IgM:Sigma) |
Millipore 05678 |
| H2BUb |
25 |
1:100 (Mab) |
1:4,000 (s.c) |
Millipore 051312 |
| Ub histones |
12-225 |
1:50 (Rb) |
1:4,000 (s.c.) |
Abcam/UbB/7780 |
| Ub histones FK1 |
12-225 |
1:100 (MsMab/IgM) |
1:9,000 (anti-IgM:Sigma) |
Enzo/FK1 PW8805 |
| H2AK5 |
14 |
1:65 (Rb) |
1:4,000 (s.c) |
Cell Signalling 2576 |
| H2BK5 |
14 |
1:200 (Rb) |
1:4,000 (s.c) |
Cell Signalling 2574 |
| H4K5 |
11 |
1:5,000 (Rb MAb) |
1:2,000 (s.c) |
Epitomics 1808-1 |
| H4K12 |
11 |
1:200 (Rb) |
1:20,000 (Sigma) |
Abcam 1761 |
| |
|
1:200 (Rb) |
1:4,000 (s.c) |
Cell Signalling 2591 |
| H4K Penta | 10 | 1:30 (Rb) | 1:20,000 (Sigma) | Millipore 06946 |
~MW is denoted as Kd. GC3α MAb is used to detect Proteasomes. Ubiquitin antibody and FK1 MAb IgM antibody are used to detect ubiquitylated and polyubiquitylated histones respectively. H4 Penta is used to detect penta-acetylated H4.