Anti-phospho-Cyclin E1 (Thr77) antibody (bs-3125R)

Anti-phospho-Cyclin E1 (Thr77) antibody

DATASHEET

Host:Rabbit

Target Protein:phospho-Cyclin E1 (Thr77)

IR:Immunogen Range:IP(p-T)PD

Clonality:Polyclonal

Isotype:IgG

Entrez Gene:898

Swiss Prot:P24864

Source:KLH conjugated Synthesised phosphopeptide derived from human Cyclin E around the phosphorylation site of Thr77:IP(p-T)PD 

Purification:affinity purified by Protein A

Storage:0.01M TBS(pH7.4) with 1% BSA, 0.03% Proclin300 and 50% Glycerol. Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles.

Background:Cyclin E is a regulatory subunit of Cdk2 and controls G1 / S transition during the mammalian cell cycle. Multiple isoforms of Cyclin E are only expressed in tumors but not in normal tissue, suggesting a post transcriptional regulation of Cyclin E. In vitro analyses indicated that these truncated variant isoforms of Cyclin E are able to phosphorylate histone H1. Alterations in the Cyclin E protein have been implicated as indicators of worse prognosis in various cancers.

Size:100ul

Concentration:1mg/ml

Applications:WB(1:500-2000)
ELISA(1:5000-10000)
Flow-Cyt(1μg /Test)

Cross Reactive Species:Human
Mouse
Rat
Chicken
Dog
Pig
Cow
Horse
Rabbit
.

For research use only. Not intended for diagnostic or therapeutic use.

VALIDATION IMAGES

Blank control:Hela.
Primary Antibody (green line): Rabbit Anti-phospho-Cyclin E1 (Thr77) antibody (bs-3152R)
Dilution: 2μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody : Goat anti-rabbit IgG-FITC
Dilution: 1μg /test.
Protocol
The cells were fixed with 4% PFA (10min 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.

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 (bs-3125R),Dilution: 3μg /10^6 cells.
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody (white blue line): Goat anti-rabbit IgG-FITC,Dilution: 1μg /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.

Blank control: MCF7.
Primary Antibody (green line): Rabbit Anti-phospho-Cyclin E1 (Thr77) antibody (bs-3125R)
Dilution: 2μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody : Goat anti-rabbit IgG-AF647
Dilution: 1μg /test.
Protocol
The cells were fixed with 4% PFA (10min 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.