PCTAIRE 1 Antibody

These findings prompted us to further explore whether CCNY-dependent CDKs could be ideal targets for breast cancer therapy. In this study, we sought to reveal a novel therapeutic target for breast cancer among CCNY-dependent CDKs by clinical correlation analysis and functional study. We found that CDK16 exhibits a distinct expression pattern in breast cancer and has strong clinical relevance, making it a potential therapeutic target for breast cancer, especially TNBC. In support of this notion, we described the oncogenic role of CDK16 in TNBC, assessed the therapeutic potential of CDK16 as a target in multiple TNBC models, and elucidated the molecular mechanisms of CDK16 involved in TNBC progression.
Succinimidyl esters should be dissolved in a solution that is free of amine-containing compounds like Tris, glycine, or protein, which will react with the SE functional group. AM esters and diacetate compounds should be dissolved in a solution that is free of serum, because serum could contain esterases that would hydrolyze the compound. Biotium guarantees the stability PCTAIRE Antibodies of chemicals, dyes, and gel stains for at least a year from the date you receive the product. However, the majority of these products are highly stable for many years, as long as they are stored as recommended. Storage conditions can be found on the product information sheet or product safety and data sheet, material safety data sheet, and on the product label.

In the validation phase, RNA was extracted from 250 μl plasma. Briefly, a 250-μl plasma aliquot was mixed with one milliliter TRIzol LS reagent. After 5 min of incubation, 266 μl of chloroform was added to the mixture. Then, the mixture was centrifuged at × g for 15 min at 4°C. An equal volume of isopropanol and 1 μl GlycoBlue was added to the solution to precipitate RNA.
Synthesized peptide derived from the Internal region of human PCTAIRE-2. Santa Cruz Biotechnology antibodies have over 360,000 research citations. Anti-PCTAIRE-3 Antibody (H-4) has 1 citations in a variety of scientific publications.

The lung metastasis was monitored by bioluminescence imaging at regular intervals. Mice were given the substrate D-luciferin (150 mg/kg) by intraperitoneal injection, then subjected to BLI using IVIS Spectrum instrument. At the experimental endpoint, the lungs were dissected, fixed and dehydrated, then the metastatic nodules on the lung surface were counted. To clearly present minor metastases, the lungs were stained with India ink, excised and decolorized before counting lung nodules.
Additional information on this brain material is detailed in Table 1. Samples were categorized based on disease stage; Non-AD , Mild Cognitive Impaired or late-AD . Tissue was homogenized in 100mM Tris-HCl (pH 7.6) containing 4% SDS, 100mM DTT and Halt protease inhibitor cocktail, heated at 100°C for 5mins, briefly sonicated and centrifuged for 15mins at 14,000 RPM. The soluble supernatant fraction was then separated from the insoluble pellet, protein concentrations were determined using Pierce 660 reagent and equal amounts of proteins were used for sample analysis by western blot.

The plasmid pBSSK-PCTAIRE-1 was kindly provided by Greg H. Enders . Sequencing of the insert revealed a single base mutation (G/A at position  727) with respect to the published sequence . PCTAIRE-1 point mutants were generated with the QuickChange site-directed mutagenesis kit and controlled by DNA sequencing. Subcellular localization of ectopically expressed PCTAIRE-1.
The results of the PDX model further verified the anti-tumor effect of targeting CDK16 in TNBC, which significantly delayed tumor occurrence (Fig. 3F), suppressed tumor growth (Fig. 3G), and finally inhibited tumor progression (Fig. 3H, I). Fresh tumor fragments from breast cancer patients were subcutaneously transplanted into 4-week female SCID/Beige mice, and the established PDX was maintained by serial passage in nude mice. Each generation was confirmed the histological and biomarker fidelity with their tumors of origin. Tumor cells were dissociated from established xenografts and infected with scramble or CDK16-shRNA lentivirus in cell suspension, then inoculated into the mammary fat pads of nude recipient mice for tumorigenesis assay.
The additional 3′ sequence contained within the PCTAIRE-3 two-hybrid clone identified here, extends exon 8 by 110 amino acids prior to an in-frame stop codon. It is possible that this represents a truncated form of PCTAIRE-3 that is normally  expressed but no such species has been detected by immunoblotting with PCTAIRE-3 antibodies. Thus,  it is probable that this represents and incompletely spliced message that was incorporated into the cDNA library. Thus, the central regions of both PCTAIRE-3a and -3b interact with Sec23Ap. We have been unable to detect any two-hybrid interaction between Sec23Ap or Sec24Dp and full-length PCTAIRE-3. GST-PCTAIRE fusion proteins (1 μg) were assayed for kinase activity by incubating immobilized GST-PCTAIRE with rat brain lysate for 30 minutes, which is required for the activation of PCTAIRE kinase activity (Graeser et al., 2002).

Cdk16 promotes tumor metastasis of mouse TNBC in systemic metastasis models. A Immunoblot analysis to verify Cdk16 knockdown in 4T1 cells and Cdk16 overexpression in EMT6 cells. B BLI images of major organs dissected from mice bearing control or Cdk16-OE EMT6 cells showed the metastatic sites of tumor cells. For lung metastasis model, MDA-MB-231-Luc cells were suspended in PBS and injected into nude recipients via lateral tail vein (5 × 105 cells/mouse for CDK16-OE analysis and 1 × 106 cells/mouse for rebastinib administration).
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Recently, rebastinib has been identified as a potent inhibitor of CDK16. It is worth noting that rebastinib is a multi-target agent against other kinases like Tie2 . We also observed the additive effect of rebastinib on Tie2-expressing macrophages in our study . Targeting CDK16 and Tie2 may jointly promote the anti-tumor activity of rebastinib in TNBC.
RNA-seq data generated in this study are publicly available in GEO dataset under accession number GSE189758. Together, these results demonstrated that pharmacological inhibition of CDK16 effectively suppresses tumor growth and metastasis of TNBC, supporting CDK16 as a promising target for the treatment of TNBC. BALB/c, Nude, and SCID/Beige mouse strains used in our study were purchased from Charles River Laboratories or Gem Pharmatech and maintained under specific-pathogen-free conditions.
Positive clones were further analysed using yeast transformation and mating protocols to determine any interaction with other COPII subunits or lamin as a negative control. Additional assays were performed using CDK2 as a negative control. A cDNA encoding amino acids of human CDK2 (terminating at the equivalent position to the PCTAIRE-3 clone isolated from screening) was amplified by PCR and cloned in-frame with the Gal4 activation domain of pGAD-T7 using standard procedures. The extended binding mode of the type II inhibitor rebastinib is quite distinct from that of the type I inhibitor, indirubin E804 .