Anti-Phospho-Cyclin D3 (Thr283) Polyclonal Antibody 2 is a Rabbit antibody targeting Phospho-Cyclin D3 (Thr283). Anti-Phospho-Cyclin D3 (Thr283) Polyclonal Antibody 2 can be used in ICC/IF,FCM.

IgG

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

1. A549 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum) at 37°C for 20 min; Antibody incubation with (phospho-Cyclin D3 (Thr283)) polyclonal Antibody, Unconjugated (TMAB-10563) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.
2. Blank control (black line): Jurkat.
Primary Antibody (green line): Rabbit Anti-phospho-Cyclin D3 (Thr283) antibody (TMAB-10563)
Dilution: 1 μg/Test;
Secondary Antibody (white blue line): Goat anti-rabbit IgG-AF488
Dilution: 0.5 μg/Test.
Isotype control (orange line): Normal Rabbit IgG
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.

ICC/IF: 1:100-500; FCM: 1μg/Test

Polyclonal

KLH conjugated synthesised phosphopeptide: human Cyclin D3 around the phosphorylation site of Thr283

Human

CCND3

G1/S-specific cyclin-D3

Regulatory component of the cyclin D3-CDK4 (DC) complex that phosphorylates and inhibits members of the retinoblastoma (RB) protein family including RB1 and regulates the cell-cycle during G(1)/S transition. Phosphorylation of RB1 allows dissociation of the transcription factor E2F from the RB/E2F complex and the subsequent transcription of E2F target genes which are responsible for the progression through the G(1) phase. Hypophosphorylates RB1 in early G(1) phase. Cyclin D-CDK4 complexes are major integrators of various mitogenenic and antimitogenic signals. Also substrate for SMAD3, phosphorylating SMAD3 in a cell-cycle-dependent manner and repressing its transcriptional activity. Component of the ternary complex, cyclin D3/CDK4/CDKN1B, required for nuclear translocation and activity of the cyclin D-CDK4 complex.