Anti-Phospho-Cyclin B1 (Ser147) Polyclonal Antibody is a Rabbit antibody targeting Phospho-Cyclin B1 (Ser147). Anti-Phospho-Cyclin B1 (Ser147) Polyclonal Antibody can be used in IHC-P,IHC-Fr,IF,FCM.

IgG

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

1. Tissue/cell: human bladder carcinoma; 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-Cyclin B1 (Ser147) Polyclonal Antibody, Unconjugated (TMAB-10560) 1: 200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining
2. Blank control: Hela.
Primary Antibody (green line): Rabbit Anti-Phospho-Cyclin B1 (Ser147) antibody (TMAB-10560)
Dilution: 1 μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody: Goat anti-rabbit IgG-AF488
Dilution: 1 μg /test.
Protocol
The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 0.1% PBST 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. Blank control (blue line): A549 (blue).
Primary Antibody (green line): Rabbit Anti-Phospho-Cyclin B1 (Ser147) antibody (TMAB-10560)
Dilution: 1 μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody (white blue line): F (ab’)2 fragment goat anti-rabbit IgG-FITC
Dilution: 1 μg /test.
Protocol
The cells were fixed with 2% paraformaldehyde (10 min) and then permeabilized with 0.1% P000 events was performed.

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

Polyclonal

KLH conjugated Synthesised phosphopeptide: human Cyclin B1 around the phosphorylation site of Ser147

Human

CCNB1

G2/mitotic-specific cyclin-B1

Essential for the control of the cell cycle at the G2/M (mitosis) transition.