Pik 75

Manufactured by Cayman Chemical

Sourced in United States, United Kingdom

PIK-75 is a laboratory reagent used for research purposes. It is a potent and selective inhibitor of the p110α isoform of phosphoinositide 3-kinase (PI3K). PIK-75 can be utilized in cell-based and biochemical assays to study the role of the p110α isoform in various biological processes.

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Lab products found in correlation

Tgx 221, by Cayman Chemical (3 mentions) As 605240, by Cayman Chemical (2 mentions) Dynasore, by Bio-Techne (1 mentions) Apc cy7, by BioLegend (1 mentions) Pi 3 4 5 p3, by Echelon Biosciences (1 mentions) Acridine orange, by Merck Group (1 mentions) Interleukin 4 (il 4), by Thermo Fisher Scientific (1 mentions) Interleukin 7 (il 7), by Thermo Fisher Scientific (1 mentions) Pe cy5, by BioLegend (1 mentions) Cyto id, by Enzo Life Sciences (1 mentions) Pacific blue conjugated anti cd3, by BioLegend (1 mentions) Il 15, by Thermo Fisher Scientific (1 mentions) Fluorescein isothiocyanate (fitc), by BioLegend (1 mentions) Cd62l, by BioLegend (1 mentions) β actin, by LI COR (1 mentions) Rapamycin, by Selleck Chemicals (1 mentions) Pi 103, by Cayman Chemical (1 mentions) Amd3100, by Selleck Chemicals (1 mentions) Anti mouse hrp secondary antibody, by Santa Cruz Biotechnology (1 mentions) Anti rabbit hrp secondary, by Abcam (1 mentions)

8 protocols using pik 75

Flow Cytometry Analysis of Immune Cells

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FITC, PE, PE-cy5, APC, APC-cy7, or Pacific Blue conjugated anti-CD3, -CD4, -CD8, -CD44, and -CD62L, were purchased from BioLegend, eBioscience, and BD Pharmingen. Anti-LC3 (PD015) and -p62 were purchased from MBL. Anti-PI(3)P, -PI(3,4)P₂, -PI(3,4,5)P₃, and PI(3,4)P₂ lipid were purchased from Echelon Biosciences. IL-7, IL-4, and IL-15 were purchased from PeproTech. Acridine orange was purchased from Sigma. CytoID was purchased from Enzo Life Sciences. PIK75 was purchased from Cayman Chemical. iSHIP (AS1949490) and Dynasore were purchased from Tocris.

McLeod I.X., Saxena R., Carico Z, & He Y.W. (2021). Class I PI3K Provide Lipid Substrate in T Cell Autophagy Through Linked Activity of Inositol Phosphatases. Frontiers in Cell and Developmental Biology, 9, 709398.

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Signaling Pathway Protein Analysis

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The primary antibodies phospho-Akt (S473), phospho-S6RP (S240/244), mTOR, raptor and rictor were all from Cell Signaling. Sin1 was from Millipore, Rheb from Bio-Rad, and β-actin was from LI-COR. The anti-mouse-HRP secondary antibody was from Santa Cruz and the anti-rabbit HRP secondary was from Abcam. The mouse-anti-CD31 used for the in vivo studies was from Dianova.
The inhibitors PI-103, PIK-75 and TGX-221 were all from Cayman Chemical. Rapamycin and AMD3100 were from Selleck Chemical and PP242 was from Chemdea. Pertussis toxin was a kind gift from the laboratory of Dr. David Fruman at UCI.

Ziegler M.E., Hatch M.M., Wu N., Muawad S.A, & Hughes C.C. (2016). mTORC2 MEDIATES CXCL12-INDUCED ANGIOGENESIS. Angiogenesis, 19(3), 359-371.

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Western Blotting Protocol for PI3K Signaling

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Antibodies used in Western immunoblotting are the following: anti-Akt (2920), -S473-Akt (9271), -pT308-Akt (C31E5E), -p110α (4249), -p110β (3011), -PARP (2730), -S6 (2217) and -pS240/S244-S6 (5364) from Cell Signaling; anti-p85α (05-212) from Millipore; anti-p85β (S3089) from Epitomics; anti-PTEN (7974) from Santa Cruz and anti-α-tubulin (T5168) from Sigma-Aldrich. Horse radish peroxidase-conjugated secondary antibodies were from Life Technologies. The PI3K selective inhibitors targeting p110α were from Selleck Chemicals (A66) and Millipore (PIK-75) and targeting p110β (TGX-221) from Cayman chemicals [34 (link), 57 (link)].

Karlsson T., Krakstad C., Tangen I.L., Hoivik E.A., Pollock P.M., Salvesen H.B, & Lewis A.E. (2016). Endometrial cancer cells exhibit high expression of p110β and its selective inhibition induces variable responses on PI3K signaling, cell survival and proliferation. Oncotarget, 8(3), 3881-3894.

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Glioblastoma Cell Culture and Signaling Modulation

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U87 and U251 human glioblastoma cell lines were cultured in DMEM (life technologies, USA) supplemented with 10% fetal bovine serum (Sigma-Aldrich), 100 units/ml penicillin-100 μg/ml streptomycin (Sigma-Aldrich). The culture was maintained at 37 °C in a humidified atmosphere containing 5% CO₂. Temozolomide (TMZ) and Ly294002 were obtained from Sigma-Aldrich. S3I-1757 was kindly provided by Dr. Said Sebti (Moffitt Cancer Center). PIK-75, TGX221, IC-87114 and AS-605240 were purchased from Cayman Chemical (USA). G418 sulfate (50 mg/ml) was obtained from Life Technologies (USA). Stat-3 siRNA was obtained from Sigma-Aldrich (USA). Rabbit anti-Human-Akt, Rabbit anti-Human phospho-Akt, Goat anti-Rabbit IgG-HRP, anti- β-actin IgG-HRP and Rabbit anti-Human cIAP2 were obtained from Santa Cruz Biotech (USA).

Wang D., Berglund A., Kenchappa R.S., Forsyth P.A., Mulé J.J, & Etame A.B. (2016). BIRC3 is a novel driver of therapeutic resistance in Glioblastoma. Scientific Reports, 6, 21710.

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BEAS-2B Cell Culture and Treatment

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BEAS-2B normal human epithelial cell line was purchased from ATCC (Catalog number: CRL-9609) and cultured according to vendor instructions using BEGM kit from LONZA (Catalog number: CC-3170). Cells were cultured on 96-well black μ-plate from ibidi (Catalog Number: 89626) for imaging studies. Tanespimycin (abcam ab141433), Acetylcysteine (Cayman, 20261), Amifostine (Cayman 14398), Bortezomib (Ayman 10008822), FK-866 (Cayman 13287), Gemcitabine (Cayman 11690), Idarubicin (Cayman 14176), NVP-AUY922 (Cayman 10012698), NVP-BEZ235 (Cayman 10565), PIK-75 (Cayman 10009210), SN-38 (Cayman 15362), Tretinoin (Cayman 11017), YM-155 (Cayman 11490), Ingenol (Cayman 14031), Sulforaphane (LKT S8044), CD-437 (Sigma C5865), and Parbendazole (Sigma 1498706) were dissolved in DMSO. 1000x concentration working solution was used for downstream experimentation.

Asano T., Chelvanambi S., Decano J.L., Whelan M.C., Aikawa E, & Aikawa M. (2022). In silico Drug Screening Approach Using L1000-Based Connectivity Map and Its Application to COVID-19. Frontiers in Cardiovascular Medicine, 9, 842641.

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Characterization of HEK293 Cells Expressing Nicotinic Receptors

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The HEK293 (293) cell line stably co-transfected with rat nAChR subunits α4 and β2 are well characterized [6 (link),11 (link),12 (link)], and they were maintained as described previously [9 (link),10 (link),14 (link)]. This cell line also expresses TNFR1 (only weak expression of TNFR2 is detected) and no additional human nAChR subunits or acetylcholine synthetic enzymes are detected [9 (link),10 (link)]. As before treatments were conducted 48 hours after cell plating [9 (link),10 (link),14 (link)]. The source and optimal concentrations of the inhibitors used are as follows: From Sigma; 1 μM nicotine (nicotine hydrogen tartrate); 250 μM choline (choline chloride); hemicholinium-3 (HC3; 50 μM). From Biosource; 25 ng/ml human TNFα. From Tocris Bioscience: 1,2,3,4,5,6-Hexabromocyclohexane (1-6Hex; 50 μM), tyrphostin 2-cyano-3-(3,4-dihydrophenyl)-N-(phenylmethyl)-2-propenamide (AG-490; dose as in text), Cucurbitacin I (100 nM); GSK690693 (5 μM), Lestaurtinib (300 nM), Ly294002 (10 μM), PI 828 (20 nM), SB2020190 (10 μM), Wortmannin (3 μM), ZM 39923 (50 μM), ZM449829 (50 μM). From Cayman Chemical: AS-605240 (20–100 nM); PIK-75 (50 μM). Most inhibitors were dissolved in either DMSO or ethanol as the carrier and in all cases the carrier was added to cultures at the same concentration as before [9 (link),10 (link),14 (link)].

Rogers S.W, & Gahring L.C. (2015). Upregulation of Nicotinic Acetylcholine Receptor alph4+beta2 through a Ligand-Independent PI3Kbeta Mechanism That Is Enhanced by TNFalpha and the Jak2/p38Mapk Pathways. PLoS ONE, 10(11), e0143319.

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Targeted Inhibition of PI3K/mTOR Signaling

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PIK-75, a p110α specific inhibitor of the PI3K complex; rapamycin, an mTOR inhibitor; and PD0325901 and PD98059, MEK inhibitors, were purchased from Cayman Chemical (Ann Arbor, MI). BKM120, a pan-PI3K inhibitor, was purchased from Active Biochem (Maplewood, NJ), and BEZ235, a dual PI3K/mTOR inhibitor, was purchased from LC Laboratories (Woburn, MA).

Valentino J.D., Li J., Zaytseva Y.Y., Mustain W.C., Elliott V.A., Kim J.T., Harris J.W., Campbell K., Weiss H., Wang C., Song J., Anthony L., Townsend CM J.r, & Evers B.M. (2014). Co-Targeting the PI3K and RAS Pathways for the Treatment of Neuroendocrine Tumors. Clinical cancer research : an official journal of the American Association for Cancer Research, 20(5), 1212-1222.

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Signaling Pathways in Platelet Activation

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Cross-linked collagen-related peptide (CRP-XL) was from R. Farndale (Department of Biochemistry, University of Cambridge, UK). Fibrillar HORM collagen (type I) derived from equine tendon from Takeda (Linz, Austria). Recombinant murine thrombopoietin (TPO) was from PeproTech (London, UK). D-phenylalanyl-L-propyl-L-arginine chloromethyl ketone (PPACK) was from Calbiochem (Merck Chemicals, Watford, UK). 3,3'-Dihexyloxacarbocyanine iodide (DiOC6) and TxB2 ELISA kit were from Enzo Life Sciences (Exeter, UK). CHRONO-LUME® reagent was from CHRONO-LOG (Labmedics, Abingdon, UK). Akt S473 (#4060), Akt T308 (#13038), Akt (#9272), ERK1/2 T202/Y204 (#9101), GSK-3α/β S21/9 (#9331), JAK2 Y1007/1008 (#3771), JAK2 (#3230), p110α (#4249), Phospho-(Ser) PKC Substrate (#2261), STAT5/ Y694 (#4322), and STAT5/ (#9358) were from Cell Signaling Technology (New England Biolabs, Hitchin, UK). GAPDH (#sc-25778) antibody was from Santa Cruz Biotechnology (Insight Biotechnology, Middlesex, UK). PD184352, VX-702 and TGX-221 were from Bio-Techne (Abingdon, UK). PIK-75 and A66 were from Cayman Chemicals (Cambridge Bioscience, Cambridge, UK). Secondary antibodies for immunoblotting were from Jackson Immunoresearch (Stratech Scientific, Ely, UK). All other reagents were from Sigma (Poole, UK), unless otherwise indicated.

Blair T.A., Moore S.F., Walsh T.G., Hutchinson J.L., Durrant T.N., Anderson K.E., Poole A.W, & Hers I. (2018). Phosphoinositide 3-kinase p110α negatively regulates thrombopoietin-mediated platelet activation and thrombus formation. Cellular signalling, 50.

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