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Polycystin 2

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Polycystin-2 is a protein that functions as a calcium-permeable cation channel. It is involved in the regulation of intracellular calcium signaling and plays a role in the development and function of various cell types.

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Lab products found in correlation

Actin, by Merck Group (2 mentions) Donkey anti goat igg hrp conjugate, by GenScript (2 mentions) P70s6k, by Santa Cruz Biotechnology (2 mentions) Polycystin 1 ct2741, by Merck Group (2 mentions) Phospho erk1 2, by Abcam (1 mentions) Phospho mtor, by Santa Cruz Biotechnology (1 mentions) Polycystin 1, by Santa Cruz Biotechnology (1 mentions) Erk1 2, by Santa Cruz Biotechnology (1 mentions) Phospho fak, by Santa Cruz Biotechnology (1 mentions) Phospho jak2, by Abcam (1 mentions) 4e bp1, by Thermo Fisher Scientific (1 mentions) Phospho p70s6k, by Santa Cruz Biotechnology (1 mentions) Goat anti mouse igg hrp conjugate, by Merck Group (1 mentions) Ap132p, by Merck Group (1 mentions) Real envision detection system, by Agilent Technologies (1 mentions) Anti pc1 7e12, by Santa Cruz Biotechnology (1 mentions) Anti pc2, by Santa Cruz Biotechnology (1 mentions) Pab2306, by Abnova (1 mentions) Actin mab1501, by Merck Group (1 mentions) Acetylated α tubulin, by Merck Group (1 mentions)

4 protocols using polycystin 2

1

Western Blot Analysis of Key Signaling Proteins

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The following primary antibodies were used for Western blot analysis: polycystin‐1 (sc‐130554, Santa Cruz Biotechnology), polycystin‐2 (sc‐10376, Santa Cruz Biotechnology), ERK1/2 (sc‐514302, Santa Cruz Biotechnology), phospho‐ERK1/2 (Abcam ab32538), mTOR (701483, Thermo Fisher Scientific), phospho‐mTOR (5536 CST), FAK (sc‐271126, Santa Cruz Biotechnology), phospho‐FAK (8556 CST), YAP (sc‐101199, Santa Cruz Biotechnology), TAZ (sc‐518026, Santa Cruz Biotechnology) and actin (MAB1501, Millipore). The following secondary antibodies were used: goat anti‐mouse IgG HRP‐conjugate (AP124P, Millipore), goat anti‐rabbit IgG HRP‐conjugate (AP132P, Millipore) and donkey anti‐goat IgG HRP‐conjugate (A00178, GenScript).
Zoi I., Gargalionis A.N., Papavassiliou K.A., Nasiri‐Ansari N., Piperi C., Basdra E.K, & Papavassiliou A.G. (2022). Polycystin‐1 and hydrostatic pressure are implicated in glioblastoma pathogenesis in vitro. Journal of Cellular and Molecular Medicine, 26(5), 1699-1709.
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2

Western Blot Analysis of Polycystic Kidney Proteins

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The following primary antibodies were used for Western blot analysis: Polycystin‐2 (sc‐10376 Santa Cruz Biotechnology), p70‐S6K (sc‐230 Santa Cruz Biotechnology), phospho‐p70‐S6K (sc‐8416 Santa Cruz Biotechnology), phospho‐mTOR (5536 CST), phospho‐4E‐BP1 (2855 CST), PTEN (9559 CST), Akt (9272 CST), phospho‐Akt (9271 CST), actin (MAB1501 Millipore), polycystin‐1 CT2741 (kindly provided by the Baltimore Polycystic Kidney Disease Research and Clinical Core Center), mTOR (701483 Thermo Fisher Scientific), 4EBP1 (AHO1382 Thermo Fisher Scientific), JAK2 (ab37226 Abcam), phospho‐JAK2 (ab32101 Abcam). The following secondary antibodies were used: goat anti‐mouse IgG HRP‐conjugate (AP124P Millipore), goat anti‐rabbit IgG HRP‐conjugate (AP132P Millipore), donkey anti‐goat IgG HRP‐conjugate (A00178 GenScript). The IgPKD1 inhibitory antibody was a generous gift from Dr O. Ibraghimov‐Beskrovnaya and H. Husson (Genzyme Co., Boston).
Papavassiliou K.A., Zoi I., Gargalionis A.N, & Koutsilieris M. (2019). Polycystin‐1 affects cancer cell behaviour and interacts with mTOR and Jak signalling pathways in cancer cell lines. Journal of Cellular and Molecular Medicine, 23(9), 6215-6227.
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3

Polycystin Signaling Pathway Regulation

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Cell culture media and all tissue culture reagents were obtained from Gibco (ThermoFisher Scientific) and Biosera. For immunohistochemistry, the following reagents were used: Dako Real Envision Detection System, peroxidase/DAB1, rabbit/mouse (Dako). The following primary antibodies were employed for Western blot analysis: polycystin‐2 (sc‐10376, Santa Cruz Biotechnology), p70S6K (sc‐230, Santa Cruz Biotechnology), phospho(p)‐p70S6K (sc‐8416, Santa Cruz Biotechnology), phospho(p)‐mTOR (5536, CST), phospho(p)‐4EBP1 (2855, CST), PTEN (9559,CST), AKT (9272,CST), phospho(p)‐AKT (9271, CST), actin (MAB1501, Millipore), polycystin‐1 CT2741 (kindly provided by the Baltimore Polycystic Kidney Disease Research and Clinical Core Center), mTOR (701483, Thermo Fisher Scientific), anti‐PC1 7E12 (sc‐130554, Santa Cruz Biotechnology) against the extracellular N‐terminal leucine‐rich domain, anti‐PC2 (sc‐28331, Santa Cruz Biotechnology) and anti‐PC2 (PAB2306, Abnova). The following secondary antibodies were used: goat anti‐mouse IgG HRP conjugate (AP124P, Millipore) and goat anti‐rabbit IgG HRP conjugate (AP132P, Millipore). The inhibitory IgPKD1 antibody (blocking the extracellular N‐terminal leucine‐rich domain) was a generous gift from Dr. O. Ibraghimov‐Beskrovnaya and H. Husson (Genzyme), and the PI3K inhibitor was obtained from Cayman, USA.
Katsianou M.A., Papavassiliou K.A., Gargalionis A.N., Agrogiannis G., Korkolopoulou P., Panagopoulos D., Themistocleous M.S., Piperi C., Basdra E.K, & Papavassiliou A.G. (2022). Polycystin‐1 regulates cell proliferation and migration through AKT/mTORC2 pathway in a human craniosynostosis cell model. Journal of Cellular and Molecular Medicine, 26(8), 2428-2437.
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4

Cell Lysis and Protein Extraction for Western Blot

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Cell lysate was prepared using a standard technique as previously described [7 (link)]. Briefly, cells were rinsed with PBS, added fresh radioimmunoprecipitation assay (Ripa) buffer containing protease inhibitor, placed on ice, gently shaken for 30 min, scraped from the culture dish, transferred to microcentrifuge tube, and spun for 15 min at 10,000 g (accuSpin Micro 17, Fisher scientific, Inc.). Total cell lysate was transferred into another microcentrifuge tube and stored in −20 °C and analyzed using a standard 6–10 % gradient sodium dodecyl sulfate—polyacrylamide gel electrophoresis. Antibody against GM3S (LSCBIO, Inc.), bicaudal C (ABGENT, Inc.), polycystin-2 (Santa Cruz, Inc.), GAPDH (cell signaling, Inc.), and acetylated-α-tubulin (Sigma, Inc.) were used at dilutions of 1:800, 1:1,000, 1:200, 1:1,000 and 1:1,000, respectively.
Mohieldin A.M., Haymour H.S., Lo S.T., AbouAlaiwi W.A., Atkinson K.F., Ward C.J., Gao M., Wessely O, & Nauli S.M. (2015). Protein composition and movements of membrane swellings associated with primary cilia. Cellular and molecular life sciences : CMLS, 72(12), 2415-2429.
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