PCTAIRE Antibody

Errors show statistical significance for the inhibition of transport following PCTAIRE-1 expression. A specific interaction was seen between Sec23Ap and the region of PCTAIRE-1 corresponding to that of PCTAIRE-3 identified in the initial screen (exons 4-8). Again, an interaction with Sec24Dp was also detectable (Fig. 1C). 1D in which the PSTAIRE kinase CDK2 is shown not to interact with any of the COPII components.
Lyse cells by adding 1X SDS sample buffer (100 µl per well of 6-well plate or 500 µl for a 10 cm diameter plate). Immediately scrape the cells off the plate and transfer the extract to a microcentrifuge tube. This product has been approved for use in this application by CST. If you have used an Abcepta PCTAIRE Antibodies product and would like to share how it has performed, please click on the "Submit Review" button and provide the requested information. Our staff will examine and post your review and contact you if needed. Provided below are standard protocols that you may find useful for product applications.

Nothing that happens in the labor other workplace is as important as your health and safety. Avantor® helps keep you safe with a robust line of safety products and personal protective equipment -- from waste bins to safety signs. If you know of any papers that use this antibody, please contact us at antibodies alzforum orgfor consideration in the References section. Immunohistochemical staining of human testis shows moderate to strong cytoplasmic positivity in cells in seminiferous ducts. Full-length recombinant human PCTK3 and Cyclin Y (2-end) were co-expressed by baculovirus in Sf9 insect cells using N-terminal GST tags.
The fact that transformed cells display high levels of PCTAIRE-1 protein suggests that the kinase is likely to be involved in proliferation. Contrary to cdk5, which is widely expressed but displays kinase activity only in neuronal cells , PCTAIRE-1 appears to be active in both normal and transformed cells . We have identified a direct interaction between the COPII machinery and a ubiquitously expressed protein kinase family, PCTAIRE. Furthermore, we have shown that manipulation of PCTAIRE kinase activity modulates membrane trafficking through the early secretory pathway. These findings are of particular interest for a number of reasons. First, there is considerable evidence for protein kinase regulation of membrane traffic at the ER-Golgi interface.

Crown antibodies pass additional stringent quality requirements, including extended control sets, uniform results against multiple biologically relevant cell lines and tissues, and function in multiple applications. RNA overexpressed in cancer tissues is not necessarily expressed at a high level in the peripheral blood. This is because the tumor mass accounts only for a small portion of the body, and both the tumor tissue and other normal tissues contribute RNAs to the circulation. Most RNAs elevated in tumor tissues are thus masked by background RNAs from normal tissues. Therefore, the RNA candidates overexpressed in tumor tissues are not necessarily good markers in the circulation.
Furthermore, CDK16 knockdown reduced tumour volume in mouse xenograft models of colorectal cancer . Interestingly, CDK16 knockdown did not affect proliferation in non-transformed cells . Taken together, these data identify CDK16 as a potential target for the development of novel anti-cancer drugs.

We next investigated the functional role of CDK16 in TNBC progression in vitro and in vivo. CDK16 was knocked down (CDK16-KD) in representative TNBC cell lines (MDA-MB-231, MDA-MB-468 and HCC1937) using two separate shRNA sequences targeting CDK16. CDK16 was efficiently knocked down, as confirmed by western blot analysis (Fig. S2A). Cells were seeded on coverslips in 24-well plates and grew for 24 h. Before imaging, coverslips were mounted using ProLong Gold Antifade Reagent with DAPI (catalog no. P36931; Thermo Fisher Scientific).
Indirect immunofluorescence revealed that the kinase is localized in the cytoplasm, and no change in the subcellular localization of PCTAIRE-1 was observed during cell cycle progression. These data argue against a role for the presumed NH2-terminal nuclear localization signal in vivo. The staining pattern shown by PCTAIRE-1 prompted us to address its potential colocalization with structures of the cytoskeletal network. Analysis of PCTAIRE-1 distribution by confocal microscopy revealed that the regions of PCTAIRE-1 staining did not correspond to particular structures of the actin and tubulin network or the endoplasmic reticulum.
Regulation of the G2/M transition in Xenopus oocytes by the cAMP-dependent protein kinase. For the indirubin E804 complex, CDK16 protein was buffered in 25 mM HEPES (pH 7.5), 250 mM NaCl, 5% glycerol and 10 mM DTT. Protein was concentrated to 15 mg/ml in the presence of the inhibitor . Crystals were grown by micro-seeding at 20°C in 130 nl of sitting drops, mixing 96 nl of protein solution with 10 nl of micro-seed solution and 24 nl of a reservoir solution containing 2.1 M Na-formate (pH 7.0), 0.1 M Bis–Tris (pH 7.0). On mounting, crystals were cryoprotected with reservoir solution mixed with 25% glycerol and vitrified in liquid nitrogen. After 24 hours, cells were treated with either 5μM Aβ42 or DMSO, which served as a vehicle control.

Further investigations will be needed to clarify the physiological functions and pathological roles of PCTK3. The binding of rebastinib to endogenous CDK16 was additionally confirmed by a CETSA, a recently developed methodology which takes advantage of alterations in protein thermal stability upon ligand binding . Aliquots of IGR-37 melanoma cells were treated with either rebastinib (20 µM) or vehicle and then heated for 3 min at different temperatures ranging from 37 to 73°C. In the DMSO-treated control cells, CDK16 was stable up to 49°C, whereas rebastinib protected CDK16 from unfolding at higher temperatures up to 57°C . We verified that the immunoreactive bands were specific using CDK16 knockdown cells as controls .
Expression of Gal4-CDK2 fusion protein was confirmed by immunoblotting . As with PCTAIRE-3, we were unable to detect a two-hybrid interaction with full-length PCTAIRE-1. Transport assays were performed essentially as described (Stephens et al., 2000). Briefly, HeLa cells were transfected with YFP-PCTAIRE-1 and ts045-G-CFP using Fugene6 . After incubation at 39.5°C for 16 hours, cells were shifted to 32°C for 60 or 90 minutes and subsequently fixed in 4% paraformaldehyde in PBS. After quenching with 30 mM glycine in PBS, cells were processed for immunofluorescence using anti-VG to quantify the amount of ts045-G-CFP delivered to the plasma membrane.

Kits often are functional for significantly longer than the guaranteed shelf life. Most of our products are stable at room temperature for many days, so in all likelihood the product will still work just fine. To be on the safe side, we recommend performing a small scale positive control experiment to confirm that the product still works for your application before processing a large number of samples or precious samples. Function May play a role in terminally differentiated neurons.
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The hydroxybutyloxime moiety extends across the expected ribose- and phosphate-binding sites of the ATP  pocket. Here, the flipped-out conformation of Phe305 in the activation segment establishes a cage-like structure around the inhibitor. The binding therefore features an induced fit that maximises the kinase–inhibitor interaction. For the rebastinib complex, CDK16 protein was buffered in 25 mM HEPES (pH 7.5), 150 mM NaCl, 5% glycerol and 10 mM DTT.