Anti-CCNE1 Polyclonal Antibody is a Rabbit antibody targeting CCNE1. Anti-CCNE1 Polyclonal Antibody can be used in WB,IHC-P,IHC-Fr,IF,FCM.

IgG

Human,Mouse,Rat

1. Tissue/cell: rat testis tissue;4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01M, pH6.0), Boiling bathing for 15 min; Blocking buffer (normal goat serum) at 37°C for 20 min;
Incubation: Anti-Cyclin E Polyclonal Antibody, Unconjugated (TMAB-00319) 1:200, overnight at 4°C; The secondary antibody was Goat Anti-Rabbit IgG, Cy3 conjugated used at 1:200 dilution for 40 minutes at 37°C.
2. Tissue/cell: human laryngocarcinoma; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01M, pH6.0), Boiling bathing for 15 min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min; Blocking buffer (normal goat serum) at 37°C for 20 min;
Incubation: Anti-Cyclin-E Polyclonal Antibody, Unconjugated (TMAB-00319) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAb staining.
3. Cell: NIH/3T3
Concentration:1:100
Host/Isotype: Rabbit/IgG
Flow cytometric analysis of primary antibody (Cat#: TMAB-00319) on NIH/3T3 (green) compared with Rabbit IgG isotype control in the absence of primary antibody (blue) followed by Alexa Fluor 488-conjugated goat anti-rabbit Igg (H+L) secondary antibody.
4. Blank control (blue line): Mouse spleen cells (blue).
Primary Antibody (green line): Rabbit Anti-Cyclin E1 antibody (TMAB-00319)
Dilution: 1 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.
Protocol
The cells were fixed with 70% ethanol (overninght at 4°C) and then permeabilized with 0.1% PBS-Tween for 20 min at room temperature. 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.
5. Blank control: MCF7. Primary Antibody (green line): Rabbit Anti-Cyclin E1 antibody (TMAB-00319)
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°C. 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.
6. Sample:
LOVO (Human) Cell Lysate at 30 μg
Thymus (Mouse) Lysate at 40 μg
U2os (Human) Cell Lysate at 30 μg
K562 (Human) Cell Lysate at 30 μg
Primary: Anti-Cyclin E1 (TMAB-00319) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 45 kDa
Observed band size: 50 kDa

WB,IHC-P,IHC-Fr,IF,FCM

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

Polyclonal

KLH conjugated synthetic peptide: rat Cyclin E

Rat

Cyclin E family,Cyclins,Cyclin E Family,Cyclin E Family