The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/500. Predicted molecular weight: 45 kDa.
Use at an assay dependent concentration.
For unpurified use at 1/100 - 1/250.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
Is unsuitable for IP.
The human Cyclin E2 gene encodes a 404 amino acid protein that is most closely related to Cyclin E. Cyclin E2 mRNA levels peaks at the G1 / S transition. Cyclin E2 associates with Cdk2 in a functional kinase complex that is inhibited by both p27 (Kip1) and p21 (Cip1). Cyclin E2 / Cdk2 phosphorylates histone H1 in vitro. G1 cyclin E controls the initiation of DNA synthesis by activating CDK2. Abnormally high levels of cyclin E expression have frequently been observed in human cancers. Unlike Cyclin E1, which is expressed in great majority of proliferating normal and neoplastically transformed cells, Cyclin E2 levels are low to undetectable in non transformed cells and increase significantly in neoplasm derived cells.
Immunocytochemistry/Immunofluorescence analysis of Jurkat cells labelling Cyclin E2 with purified ab32103 at 1/2000. Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% tritonX-100. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (ab150077) at 1/1000 dilution was used as the secondary antibody. Nuclei counterstained with DAPI (blue).
Secondary Only Control: PBS was used instead of the primary antibody as the negative control.
Overlay histogram showing HeLa cells stained with ab32103 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32103, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
Immunocytochemistry/ Immunofluorescence - Anti-Cyclin E2 antibody [E142] (ab32103)This image is courtesy of an abreview by Kirk Mcmanus
ab32103 staining Cyclin E2 in the Hela cell line from Human cervix by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.5% Triton X-100. Samples were incubated with primary antibody (1/500 in PBS) for 1 hour at 22°C. Ab150081 (1/200) was used as the secondary antibody.
Vannini I et al. Transcribed ultraconserved region 339 promotes carcinogenesis by modulating tumor suppressor microRNAs. Nat Commun8:1801 (2017).
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Vogel KU et al. The RNA-Binding Proteins Zfp36l1 and Zfp36l2 Enforce the Thymic ß-Selection Checkpoint by Limiting DNA Damage Response Signaling and Cell Cycle Progression. J Immunol197:2673-85 (2016).
Read more (PubMed: 27566829) »