Source and reaction conditions of antibodies for Western blot analysis
Target . | Antibody source (antibody) . | Dilution of primary antibody; blocking . | Size of protein . |
|---|---|---|---|
| AML-1 | OR (Ab-2) | 1:40; 5% milk | 53-55 kDa78 |
| c-myb | SC (H-141) | 1:1000; 3% BSA | 75 kDa |
| C/EBP-α | SC (14-AA) | 1:1000; 3% BSA | 42 and 30 kDa |
| C/EBP-β | SC (C-19) | 1:1000; 3% BSA | 43 and 20 kDa48 |
| C/EBP-δ | SC (C-22) | 1:1000; 3% BSA | 33 kDa79 |
| C/EBP-ϵ | SC (C-22) Koeffler and colleagues80 * | 1:1000; 3% BSA | 32, 27, and 14 kDa43,80 |
| C/EBP-γ | Koeffler and colleagues83 † | 1:1000; 5% milk | 18-20 kDa66 |
| C/EBP-ζ | SC (B-3) | 1:1000; 3% BSA | 29 kDa81 |
| CDP | Koeffler and colleagues58 ‡ (α-861) | 1:1000; 2.5% BSA, 5% milk | 160-180 kDa82 |
| c-fos | BD (G54-9.9) | 1:1000; 5% BSA | 55, 57, 60, and 62 kDa |
| c-jun | BD (clone 3) | 1:1000; 3% BSA | 39 kDa |
| PU.1 | SC (T-21) | 1:1000; 3% BSA | 40 kDa |
| Elf-1 | SC (C-20) | 1:1000; 2.5% milk | ≈95 kDa, as a double band |
| α-tubulin | SA (DM 1A) | 1:4500; 3% BSA | 55 kDa |
Target . | Antibody source (antibody) . | Dilution of primary antibody; blocking . | Size of protein . |
|---|---|---|---|
| AML-1 | OR (Ab-2) | 1:40; 5% milk | 53-55 kDa78 |
| c-myb | SC (H-141) | 1:1000; 3% BSA | 75 kDa |
| C/EBP-α | SC (14-AA) | 1:1000; 3% BSA | 42 and 30 kDa |
| C/EBP-β | SC (C-19) | 1:1000; 3% BSA | 43 and 20 kDa48 |
| C/EBP-δ | SC (C-22) | 1:1000; 3% BSA | 33 kDa79 |
| C/EBP-ϵ | SC (C-22) Koeffler and colleagues80 * | 1:1000; 3% BSA | 32, 27, and 14 kDa43,80 |
| C/EBP-γ | Koeffler and colleagues83 † | 1:1000; 5% milk | 18-20 kDa66 |
| C/EBP-ζ | SC (B-3) | 1:1000; 3% BSA | 29 kDa81 |
| CDP | Koeffler and colleagues58 ‡ (α-861) | 1:1000; 2.5% BSA, 5% milk | 160-180 kDa82 |
| c-fos | BD (G54-9.9) | 1:1000; 5% BSA | 55, 57, 60, and 62 kDa |
| c-jun | BD (clone 3) | 1:1000; 3% BSA | 39 kDa |
| PU.1 | SC (T-21) | 1:1000; 3% BSA | 40 kDa |
| Elf-1 | SC (C-20) | 1:1000; 2.5% milk | ≈95 kDa, as a double band |
| α-tubulin | SA (DM 1A) | 1:4500; 3% BSA | 55 kDa |
The membranes were blocked in PBS with 0.5% Tween with the indicated amount of BSA and/or skimmed milk prior to addition of primary antibody. Protein sizes are as reported on the data sheets of the antibodies or, where indicated, in publications.
OR indicates Oncogene Research (Boston, MA); SC, Santa Cruz Biotechnology (Santa Cruz, CA), BD, Becton Dickinson; and SA, Sigma-Aldrich.
University of California—Los Angeles (UCLA) School of Medicine.
National Cancer Institute (NCI)—Frederick, MD.
McGill University, Montreal, QC, Canada.