This antibody gave a positive signal in Human Small Intestine tissue lysate.
This antibody gave a positive result when used in the following methanol fixed cell lines: HeLa.
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The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration.
Use a concentration of 5 µg/ml.
Use a concentration of 1 µg/ml. 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.
ICC/IF image of ab112580 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab112580 at 5µg/ml overnight at +4°C. The secondary antibody (green) was goat anti-rabbit DyLight® 488 (ab96899) IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Western blot - Anti-E2F1 antibody (ab112580)
All lanes : Anti-E2F1 antibody (ab112580) at 1 µg/ml
Lane 1 : Human small intestine tissue lysate - total protein (ab29276) Lane 2 : Human small intestine tissue lysate - total protein (ab29276) with Immunizing peptide at 1 µg/ml
Lysates/proteins at 20 µg per lane.
Secondary Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution Developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 47 kDa Observed band size : 47 kDa Additional bands at : 53 kDa. We are unsure as to the identity of these extra bands.