Anti-Phospho-p107 (Thr369) Polyclonal Antibody 2 is a Rabbit antibody targeting Phospho-p107 (Thr369). Anti-Phospho-p107 (Thr369) Polyclonal Antibody 2 can be used in WB,IHC-P,IHC-Fr,IF,FCM.

IgG

Human,Mouse,Rat (predicted:Chicken,Dog,Pig,Cow,Horse,Rabbit)

1. Blank control: Molt4.
Primary Antibody (green line): Rabbit Anti-phospho-p107 (Thr369) antibody (TMAB-11075)
Dilution: 2 μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody: Goat anti-rabbit IgG-APC
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-20℃. 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.
2. Sample:
Lane 1: Mouse Testis tissue lysates
Lane 2: Rat Testis tissue lysates
Lane 3: Human HeLa cell lysates
Lane 4: Human Jurkat cell lysates
Lane 5: Human 293T cell lysates
Lane 6: Human U-2 OS cell lysates
Primary: Anti-phospho-p107 (Thr369) (TMAB-11075) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 121 kDa
Observed band size: 130 kDa
3. Paraformaldehyde-fixed, paraffin embedded Mouse Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with phospho-p107 (Thr369) Polyclonal Antibody, Unconjugated (TMAB-11075) at 1: 200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.

WB: 1:500-2000; IHC-P: 1:100-500; IHC-Fr: 1:100-500; IF: 1:100-500; FCM: 2μg/Test

Polyclonal

KLH conjugated synthesised phosphopeptide: human RBL1 around the phosphorylation site of Thr369

Human

RBL1

Retinoblastoma-like protein 1

Key regulator of entry into cell division. Directly involved in heterochromatin formation by maintaining overall chromatin structure and, in particular, that of constitutive heterochromatin by stabilizing histone methylation. Recruits and targets histone methyltransferases SUV420H1 and SUV420H2, leading to epigenetic transcriptional repression. Controls histone H4 'Lys-20' trimethylation. Probably acts as a transcription repressor by recruiting chromatin-modifying enzymes to promoters. Potent inhibitor of E2F-mediated trans-activation. Forms a complex with adenovirus E1A and with SV40 large T antigen. May bind and modulate functionally certain cellular proteins with which T and E1A compete for pocket binding. May act as a tumor suppressor.