Anti-Phospho-E2F1 (Ser332) Polyclonal Antibody is a Rabbit antibody targeting Phospho-E2F1 (Ser332). Anti-Phospho-E2F1 (Ser332) Polyclonal Antibody can be used in WB,IHC-P,IHC-Fr,IF,FCM.

IgG

Human,Mouse,Rat

1. Paraformaldehyde-fixed, paraffin embedded (rat colon); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (phospho-E2F1 (Ser332)) Polyclonal Antibody, Unconjugated (TMAB-10610) at 1: 200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
2. Blank control (Black line): Molt4 (Black).
Primary Antibody (green line): Rabbit Anti-phospho-E2F1 (Ser332) antibody (TMAB-10610)
Dilution: 1 μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody (white blue line): Goat anti-rabbit IgG-AF647
Dilution: 1 μg /test.
Protocol
The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at room temperature. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.
3. Tissue/cell: mouse intestine tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01 M, pH 6.0), Boiling bathing for 15 min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min; Blocking buffer (normal goat serum) at 37℃ for 20 min;
Incubation: Anti-phospho-E2F1 (Ser332) Polyclonal Antibody, Unconjugated (TMAB-10610) 1: 200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining
4. Tissue/cell: human colon cancer; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01 M, pH 6.0), Boiling bathing for 15 min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min; Blocking buffer (normal goat serum) at 37℃ for 20 min;
Incubation: Anti-phospho-E2F1 (Ser332) Polyclonal Antibody, Unconjugated (TMAB-10610) 1:500, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining
5. Tissue/cell: human skin tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01 M, pH 6.0), Boiling bathing for 15 min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min; Blocking buffer (normal goat serum) at 37℃ for 20 min;
Incubation: Anti-phospho-E2F1 (Ser332) Polyclonal Antibody, Unconjugated (TMAB-10610) 1:500, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining
6. Sample:
Lane 1: Human Hela cell lysates
Lane 2: Human A673 cell lysates
Lane 3: Human Molt-4 cell lysates
Lane 4: Human U251cell lysates
Lane 5: Human HepG2 cell lysates
Primary: Anti-phospho-E2F1 (Ser332) (TMAB-10610) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 46 kDa
Observed band size: 61 kDa

WB: 1:500-2000; IHC-P: 1:100-500; IHC-Fr: 1:100-500; IF: 1:100-500; FCM: 1μg/Test

Polyclonal

KLH conjugated Synthesised phosphopeptide: human E2F1 around the phosphorylation site of Ser332

Human

E2F1

Transcription factor E2F1

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.