PCTAIRE Antibodies

PCTAIRE-1 activity is cell cycle dependent and displays a peak in the S and G2 phases. We show that the low level of kinase activity observed until the onset of S phase correlates with elevated tyrosine phosphorylation of the molecule. CDK16 is highly expressed in breast cancer, particularly in triple-negative breast cancer . The elevated CDK16 expression is correlated with poor outcomes in breast cancer patients.
Heat map analysis showed that the target genes of the Rb-E2F pathway were down-regulated upon Reb treatment (Fig. 7E) or CDK16 knockdown (Fig. 7F). Immunoblot analysis validated that Rb phosphorylation was significantly decreased upon Reb treatment PCTAIRE Antibodies (Fig. 7G) or CDK16 knockdown (Fig. 7H). In addition, the expression of genes directly regulated by Rb-E2F signal was significantly down-regulated upon Reb treatment (Fig. 7I) or CDK16 knockdown (Fig. 7J), as measured by qPCR analysis.

We found that the extent of PCTAIRE-1 activation inversely correlates with the content in phosphotyrosine, which is high in G0 and during progression through G1. As observed for other cdk family members, the PCTAIRE-1 level did not change during transition through the  cell division cycle. The catalytic domain of PCTAIRE-1 contains all of the conserved features of the Ser/Thr protein kinase family. To test whether PCTAIRE-1 displays phosphotransferase activity, we immunoprecipitated HA-tagged PCTAIRE-1 and used MBP as an acceptor substrate. HA-tagged PCTAIRE-1 appeared to efficiently phosphorylate MBP (Fig. 2,D, Lane 3).
Abcepta validation program described in the recent antibody quality review published in the May 2015 issue of Nature. Diagram of the study design, clinical sample flow, and process of candidate gene evaluation. The aim of this study was to identify circulating mRNA markers of NSCLC. The study design included a discovery phase and a validation phase. The whole experimental strategy is simplified in a flow chart in Figure 1.

Crissman H. A., Hirons G. T. Staining of DNA in live and fixed cells Darnzynkiewicz Z. Robinson J. P. Crissman H. A. Kozak M. An analysis of 5′-noncoding sequences from 699 vertebrate messengers RNAs. Blangy A., Lane H. A., d’Herdin P., Harper M., Kress M., Nigg E. A. Phosphorylation by p34cdc2 regulates spindle association of human Eg5, a kinesin-related motor essential for bipolar spindle formation in vivo. Ziebold U., Bartsch O., Marais R., Ferrari S., Klempnauer K. H. Phosphorylation and activation of B-Myb by cyclin A-Cdk2. Harper J. W., Elledge S. J. The role of Cdk7 in CAK function, a retro-retrospective. Morley S. J. Regulation of components of the translational machinery by protein phosphorylation Clemens M.
Clinical kinase inhibitors were obtained from the FIMM drug collection. Published kinase inhibitor set compounds were a gift from William Zuercher . Unless otherwise indicated, all other reagents were obtained from Sigma. Okuda T., Cleveland J. L., Downing J. R. PCTAIRE-1 and PCTAIRE-3, two members of a novel cdc2/CDC28-related protein kinase gene family. Lee M. G., Nurse P. Complementation used to clone a human homologue of the fission yeast cell cycle control gene cdc2.

S-phase extracts treated with no antibody or immunoprecipitated with anti-cyclin A antibody were used as negative and positive controls, respectively. The results of an experiment representative of three independent determinations are shown. C, an aliquot (15 μg) of the extracts used in A was probed with PCTAIRE-1 antibody to examine the level of endogenous PCTAIRE-1 expression during cell cycle progression. D, extracts (200 μg) made in G1 or upon release from the hydroxyurea block (see “Materials and Methods”) were immunoprecipitated using a monoclonal antiphosphotyrosine antibody.
Find CDK16 Antibodies validated for a specific application such as WB, ELISA, IF, IHC. Some of the available applications are listed below. One-stop service from gene synthesis and vector construction to protein expression and purification. A- and B-type cyclins differentially modulate substrate specificity of cyclin-cdk complexes. The structural basis for specificity of substrate and recruitment peptides for cyclin-dependent kinases.
Gao C. Y., Chauthaiwale V. M., Rampalli A. M., Zelenka P. S. Expression of alternatively spliced PCTAIRE-1 mRNA in PC12 cells and neonatal rat brain. Sauer K., Lehner C. F. The role of cyclin E in the regulation of entry into S phase. Tris (pH 6.8), 10% glycerol, 2% SDS, 0.2% bromphenol blue, and 0.1 M DTT]. Proteins were resolved on an 8% Laemmli gel, transferred to PVDF membrane, and stained with 0.1% Ponceau S solution. Western blot with anti-PCTAIRE-1 antibody was performed as described above. This website is using a security service to protect itself from online attacks.

PCTAIRE antibody LS-C is an unconjugated rabbit polyclonal antibody to PCTAIRE (N-Terminus) from human. Western blot of lysates performed using standard western blot reagents and 4-20% SDS-PAGE. Hirose T., Tamaru T., Okumura N., Nagai K., Okada M. PCTAIRE-2, a Cdc2-related serine/threonine kinase, is predominantly expressed in terminally differentiated neurons. Peeper D. S., Parker L. L., Ewen M. E., Toebes M., Xu M., Zantema A., van der Eb A. J., Piwnica-Worms H. A- and B-type cyclins differentially modulate substrate specificity of cyclin-cdk complexes. Tsai L. H, Takahashi, T., Caviness V. S., Jr., Harlow E. Activity and expression pattern of cyclin-dependent kinase 5 in the embryonic mouse nervous system.
Despite identifying an interaction between Sec31Ap and the truncated PCTAIRE-3 clone in two hybrid-assays, we have been unable to specifically co-immunoprecipitate these two proteins . We next tested the ability of the compounds to inhibit CDK16 activity in intact cells. It was shown recently that Ser336 phosphorylation on cyclin Y is dependent on CDK16 kinase activity . We therefore tested whether inhibitor treatment would affect Ser336 phosphorylation using a phospho-specific antibody. HA-tagged cyclin Y WT or S336A mutant was co-expressed with FLAG-tagged CDK16 WT or D304A (kinase-inactive) mutant in COS1 cells. The cells were treated with increasing concentrations of the respective inhibitors for 1 h, and lysates were immunoblotted for detection of Ser336–cyclin Y phosphorylation.

All the reagents should be mixed thoroughly by gently swirling before pipetting. Keep appropriate numbers of strips for 1 experiment and remove extra strips from microtiter plate. Removed strips should be resealed and stored at -20°C until the kits expiry date. Prepare all reagents, working standards and samples as directed in the previous sections. If values for these are not within the range of the standard curve, users must determine the optimal sample dilutions for their experiments. Functional regulation of transient receptor potential canonical 7 by cGMP-dependent protein kinase Iα.
This would also explain the lack of interaction observed in the two-hybrid system between Sec23Ap and full-length Sec23Ap. We have only been able to detect an interaction with Sec31Ap in some two-hybrid assays and not in any other binding assays tested. We cannot rule out some role for this interaction, possibly in  the recruitment of PCTAIRE kinases to fully coated COPII vesicles but it seems more likely that PCTAIRE kinases do not interact with Sec31Ap.
Interestingly, CDK16 inhibition also inhibits Rb phosphorylation and Rb-E2F signal. In conclusion, our study provides new insights into the roles of atypical CDKs in cancer and emphasizes that CDK16 is a promising therapeutic target for TNBC. Publicly available breast cancer datasets analyses and Kaplan-Meier survival analyses were performed to reveal the expression and clinical relevance of atypical CDKs in breast cancer. CDK16 protein expression was further examined by immunohistochemical and immunoblot analyses of clinical samples.