The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration.
1/250. Detects a band of approximately 47 kDa (predicted molecular weight: 47 kDa).
Transcription activator that binds DNA cooperatively with dp proteins through the E2 recognition site, 5'-TTTC[CG]CGC-3' found in the promoter region of a number of genes whose products are involved in cell cycle regulation or in DNA replication. The DRTF1/E2F complex functions in the control of cell-cycle progression from G1 to S phase. E2F-1 binds preferentially RB1 protein, in a cell-cycle dependent manner. It can mediate both cell proliferation and p53-dependent apoptosis.
Belongs to the E2F/DP family.
Phosphorylated by CDK2 and cyclin A-CDK2 in the S-phase. Acetylation stimulates DNA-binding. Enhanced under stress conditions such as DNA damage and inhibited by retinoblastoma protein pRB. Regulated by KAP1/TRIM28 which recruits HDAC1 to E2F1 resulting in deacetylation. Acetylated by P/CAF/KAT2B.
Western blot - Anti-E2F1 (phospho S364) antibody (ab5391)
All lanes : Anti-E2F1 (phospho S364) antibody (ab5391) at 1/250 dilution
Lane 1 : CRE (mouse) cells transfected with human E2F1 Lane 2 : CRE (mouse) cells transfected with human E2F1 with doxorubicin Lane 3 : NiH3T3 (mouse) cell lysate Lane 4 : NiH3T3 (mouse) cell lysate with doxorubicin Lane 5 : MDA-231 (human breast cancer) cell lysate Lane 6 : MDA-231 (human breast cancer) cell lysate with doxorubicin
Secondary All lanes : HRP conjugated Goat-anti-Rabbit IgG MX10
Predicted band size: 47 kDa
Lysates were prepared from untreated cells and cells treated with 2µM Doxorubicin used as a DNA damaging agent. The arrow indicates E2F1.
Phosphorylated E2F1 was only expressed in the treated cells. The blot clearly shows that ab5391 only detected phosphorylated E2F1 in the human cell lines.