Anti-Phospho-Cyclin E1 (Thr77) Polyclonal Antibody is a Rabbit antibody targeting Phospho-Cyclin E1 (Thr77). Anti-Phospho-Cyclin E1 (Thr77) Polyclonal Antibody can be used in FCM.

IgG

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

1. Blank control (blue line): MCF7 (fixed with 70% ethanol (Overnight at 4℃) and then permeabilized with 90% ice-cold methanol for 30 min on ice)
Primary Antibody (green line): Rabbit Anti-phospho-Cyclin E1 (Thr77) antibody (TMAB-10564), Dilution: 3 μg /10^6 cells.
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody (white blue line): Goat anti-rabbit IgG-FITC, Dilution: 1 μg /test.
The cells werE. Cells stained with Primary Antibody for 30 min at room temperature. The cells were then incubated in 1 X PBS/2%BSA/10% goat serum to block non-specific protein-protein interactions followed by the antibody for 15 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.
2. Blank control: Hela.
Primary Antibody (green line): Rabbit Anti-phospho-Cyclin E1 (Thr77) antibody
Dilution: 2 μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody: Goat anti-rabbit IgG-FITC
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: MCF7. Primary Antibody (green line): Rabbit Anti-phospho-Cyclin E1 (Thr77) antibody (TMAB-10564) Dilution: 2 μg /10^6 cells; isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody: 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-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.

FCM: 1μg/Test

Polyclonal

KLH conjugated Synthesised phosphopeptide: human Cyclin E around the phosphorylation site of Thr77

Human

CCNE1

G1/S-specific cyclin-E1

Essential for the control of the cell cycle at the G1/S (start) transition.