PCTAIRE Antibodies
Interactions of indirubin E804 and rebastinib with the ATP-binding pocket of CDK16. Share with us your publications using BT LAB product and get free products. Peptides were analyzed on a Q-Exactive Plus following with a 75µm x 50cm C18 reversed-phase-UPLC column using a 90 minute gradient on an EASY-nLC 1000 system .
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.
MDA-MB-231 cells with CDK16-KD were orthotopically implanted into the mammary fat pads of nude recipients, and MDA-MB-468 cells with CDK16-KD were subcutaneously inoculated into the same strain recipients for tumorigenesis assay. Consistent results were obtained from the MDA-MB-468 xenograft model (Fig. 2J-M). Together, these results demonstrated that CDK16 is critical for TNBC progression.
The newly proposed atypical CDK members PFTAIRE1–2 (CDK14–15) and PCTAIRE1–3 (CDK16–18) kinases share CCNY as an activator and are co-expressed in various tissues with the highest expression in brain and testis . The functions and regulatory networks of these atypical CDKs are largely unknown. Knocking down individual atypical CDKs failed to cause any phenotype, suggesting possible functional redundancy of these kinases . However, a functional study of CDK16 using CDK16 conditional knockout mouse model has validated its essential role in spermatogenesis . Here, our study demonstrated that CDK16 phosphorylates PRC1 and Rb, thereby regulating spindle formation and the Rb-E2F pathway, providing new insights into the function of the atypical CDK subfamily.
RNAi depletion of PCTAIRE-1 caused dispersion of ERGIC-53 from a more punctate localization in lamin siRNA transfected cells to a more ER-like pattern in PCTAIRE-1 siRNA-transfected cells Fig. An indistinguishable phenotype was seen with two independent siRNA duplexes targeting PCTAIRE-1 . The localization of Sec24Dp in siRNA-transfected cells was also found to be more dispersed in these cells, although consistent, this is in fact a very mild phenotype. Recombinant FLAG-Sec23Ap was expressed in insect cells using a baculovirus expression plasmid kindly provided by Peter Espenshade and Sf9 cells grown in ExCell 420 medium . FLAG-Sec23Ap was affinity-purified using anti-FLAG M2 antibody immobilized on Sepharose beads.
Immunoprecipitated proteins were then detected by immunoblotting using anti-Sec23Ap or anti-PCTAIRE-1/3 antibodies . The crystal structures of monomeric CDK16 represent the first structures from the PCTAIRE family of CDK kinases . The kinase domain exhibits the classical bilobal architecture intermixed with short insertions that help to define the CDK family fold. The complex with indirubin E804 displays some disorder in the kinase N-lobe, consistent with its requirement for a cyclin partner.
It is possible that PCTAIRE activity has a role in the organization of ERES in polarizing cells or in the selective export of specific cargo proteins that are required for directed cell migration and/or polarization. Clearly, the roles of this and other protein phosphorylation events in the regulation of transport through the secretory pathway require further investigation. PCTAIRE-1 migrated in SDS polyacrylamide gels with an apparent Mr of 55,000, in agreement with the predicted size of the polypeptide .
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The membrane was probed with PCTAIRE-1 antibody or with the same antibody preincubated with a 10-fold (w/w) excess of antigenic peptide . D and E, COS-7 cells ectopically expressing HA-tagged PCTAIRE-1 were extracted at 24 or 36 h after transfection. Controls include nontransfected cells and cells transfected with empty vector (pcDNA3-HA; Lane 2). PCTAIRE-1 protein was revealed with the anti-PCTAIRE-1 antibody or with the anti-HA 12CA5 antibody .
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.
These particles are protected by the plasma membrane and are considered the major sources of circulating RNA. This notion is also confirmed by the observation that nude RNA is very unstable in the blood, which is rich in RNase activity . Information about which particle PCTK1 mRNA is derived from will be helpful in the development of better diagnostic methods. For example, different methods have been developed to capture extracellular vesicles . If PCTK1 mRNA exists in the captured vesicles, these technologies can be applied in the enrichment of circulating PCTK1 RNA in the detection procedure.
PCTAIRE kinases are almost twice the size of other members of the family because of NH2- and COOH-terminal extensions. Some members of the cdk family are predominantly expressed in terminally differentiated cells , including PCTAIRE-2 . To investigate the role played by PCTAIRE-1 in the cell division cycle, we began by analyzing the kinase’s pattern of protein expression. The data obtained indicated that contrary to PCTAIRE-2 , PCTAIRE-1 is expressed across the entire panel of cell lines that we have examined.
Cells were maintained in appropriate medium supplemented with 10% fetal bovine serum (PAN-Biotech) and 1% penicillin/streptomycin at 37 °C incubators with 5% CO2. See the product information sheets for specific reactive dyes more information. The information contained on this Web Site is provided for informational purposes only.
In this study, we demonstrated that CDK16 is a critical oncogenic driver in breast cancer. CDK16 is up-regulated in breast cancer with aberrantly high levels in TNBC, and elevated CDK16 expression is correlated with poor prognosis of breast cancer patients. Knockdown of CDK16 significantly inhibits the proliferation of TNBC and HR+ breast cancer cells, but only has a slight effect on HER2+ breast cancer cells. Either genetic depletion or pharmacological inhibition of CDK16 effectively suppresses tumor growth and metastasis of TNBC in various tumor models.
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.
MDA-MB-231 cells with CDK16-KD were orthotopically implanted into the mammary fat pads of nude recipients, and MDA-MB-468 cells with CDK16-KD were subcutaneously inoculated into the same strain recipients for tumorigenesis assay. Consistent results were obtained from the MDA-MB-468 xenograft model (Fig. 2J-M). Together, these results demonstrated that CDK16 is critical for TNBC progression.
The newly proposed atypical CDK members PFTAIRE1–2 (CDK14–15) and PCTAIRE1–3 (CDK16–18) kinases share CCNY as an activator and are co-expressed in various tissues with the highest expression in brain and testis . The functions and regulatory networks of these atypical CDKs are largely unknown. Knocking down individual atypical CDKs failed to cause any phenotype, suggesting possible functional redundancy of these kinases . However, a functional study of CDK16 using CDK16 conditional knockout mouse model has validated its essential role in spermatogenesis . Here, our study demonstrated that CDK16 phosphorylates PRC1 and Rb, thereby regulating spindle formation and the Rb-E2F pathway, providing new insights into the function of the atypical CDK subfamily.
RNAi depletion of PCTAIRE-1 caused dispersion of ERGIC-53 from a more punctate localization in lamin siRNA transfected cells to a more ER-like pattern in PCTAIRE-1 siRNA-transfected cells Fig. An indistinguishable phenotype was seen with two independent siRNA duplexes targeting PCTAIRE-1 . The localization of Sec24Dp in siRNA-transfected cells was also found to be more dispersed in these cells, although consistent, this is in fact a very mild phenotype. Recombinant FLAG-Sec23Ap was expressed in insect cells using a baculovirus expression plasmid kindly provided by Peter Espenshade and Sf9 cells grown in ExCell 420 medium . FLAG-Sec23Ap was affinity-purified using anti-FLAG M2 antibody immobilized on Sepharose beads.
Immunoprecipitated proteins were then detected by immunoblotting using anti-Sec23Ap or anti-PCTAIRE-1/3 antibodies . The crystal structures of monomeric CDK16 represent the first structures from the PCTAIRE family of CDK kinases . The kinase domain exhibits the classical bilobal architecture intermixed with short insertions that help to define the CDK family fold. The complex with indirubin E804 displays some disorder in the kinase N-lobe, consistent with its requirement for a cyclin partner.
It is possible that PCTAIRE activity has a role in the organization of ERES in polarizing cells or in the selective export of specific cargo proteins that are required for directed cell migration and/or polarization. Clearly, the roles of this and other protein phosphorylation events in the regulation of transport through the secretory pathway require further investigation. PCTAIRE-1 migrated in SDS polyacrylamide gels with an apparent Mr of 55,000, in agreement with the predicted size of the polypeptide .
We found alternative products that are currently in stock . To compare product details, select up to 3 alternatives below and click Compare Selected. To replace the original item in your cart, enter a quantity and click Replace in Cart to swap products. These Terms and Conditions of Use are applicable to users in the United States, Canada, and Puerto Rico.
The membrane was probed with PCTAIRE-1 antibody or with the same antibody preincubated with a 10-fold (w/w) excess of antigenic peptide . D and E, COS-7 cells ectopically expressing HA-tagged PCTAIRE-1 were extracted at 24 or 36 h after transfection. Controls include nontransfected cells and cells transfected with empty vector (pcDNA3-HA; Lane 2). PCTAIRE-1 protein was revealed with the anti-PCTAIRE-1 antibody or with the anti-HA 12CA5 antibody .
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.
These particles are protected by the plasma membrane and are considered the major sources of circulating RNA. This notion is also confirmed by the observation that nude RNA is very unstable in the blood, which is rich in RNase activity . Information about which particle PCTK1 mRNA is derived from will be helpful in the development of better diagnostic methods. For example, different methods have been developed to capture extracellular vesicles . If PCTK1 mRNA exists in the captured vesicles, these technologies can be applied in the enrichment of circulating PCTK1 RNA in the detection procedure.
PCTAIRE kinases are almost twice the size of other members of the family because of NH2- and COOH-terminal extensions. Some members of the cdk family are predominantly expressed in terminally differentiated cells , including PCTAIRE-2 . To investigate the role played by PCTAIRE-1 in the cell division cycle, we began by analyzing the kinase’s pattern of protein expression. The data obtained indicated that contrary to PCTAIRE-2 , PCTAIRE-1 is expressed across the entire panel of cell lines that we have examined.
Cells were maintained in appropriate medium supplemented with 10% fetal bovine serum (PAN-Biotech) and 1% penicillin/streptomycin at 37 °C incubators with 5% CO2. See the product information sheets for specific reactive dyes more information. The information contained on this Web Site is provided for informational purposes only.
In this study, we demonstrated that CDK16 is a critical oncogenic driver in breast cancer. CDK16 is up-regulated in breast cancer with aberrantly high levels in TNBC, and elevated CDK16 expression is correlated with poor prognosis of breast cancer patients. Knockdown of CDK16 significantly inhibits the proliferation of TNBC and HR+ breast cancer cells, but only has a slight effect on HER2+ breast cancer cells. Either genetic depletion or pharmacological inhibition of CDK16 effectively suppresses tumor growth and metastasis of TNBC in various tumor models.
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