Mouse, Human Predicted to work with:
Rat, Chinese hamster
Synthetic peptide corresponding to Human Cyclin E1 aa 50-150 conjugated to Keyhole Limpet Haemocyanin (KLH). (Peptide available as ab101079, ab101079)
This antibody gave a positive signal in HepG2 whole cell lysate and in the following mouse tissue lysates: Brain; Testis; Heart.
This antibody gave a positive result in IHC in the following FFPE tissue: Human normal placenta.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 µg/ml. Detects a band of approximately 47 kDa (predicted molecular weight: 47 kDa).
Use a concentration of 1 µg/ml.
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
FunctionEssential for the control of the cell cycle at the G1/S (start) transition.
Tissue specificityHighly expressed in testis and placenta. Low levels in bronchial epithelial cells.
Sequence similaritiesBelongs to the cyclin family. Cyclin E subfamily.
Post-translational modificationsPhosphorylation of Thr-395 by GSK3 and of Ser-399 by CDK2 accelerates degradation via the ubiquitin proteasome pathway. Phosphorylated upon DNA damage, probably by ATM or ATR.
IHC image of Cyclin E staining in Human normal placenta formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab88259, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
Western blot - Anti-Cyclin E1 antibody (ab88259)
All lanes : Anti-Cyclin E1 antibody (ab88259) at 1 µg/ml
Lane 1 : Brain (Mouse) Tissue Lysate Lane 2 : Testis (Mouse) Tissue Lysate Lane 3 : Heart (Mouse) Tissue Lysate Lane 4 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution Developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 47 kDa Observed band size : 47 kDa Additional bands at : 32 kDa,65 kDa,70 kDa. We are unsure as to the identity of these extra bands.
ICC/IF image of ab88259 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab88259, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) Hek293, HepG2 and MCF7 cells at 1µg/ml.
References for Anti-Cyclin E1 antibody (ab88259)
This product has been referenced in:
Bonazzi VF et al. Cross-Platform Array Screening Identifies COL1A2, THBS1, TNFRSF10D and UCHL1 as Genes Frequently Silenced by Methylation in Melanoma. PLoS One6:e26121 (2011).
Read more (PubMed: 22028813) »