Pim447

Manufactured by Selleck Chemicals

Sourced in Italy

PIM447 is a laboratory equipment product designed for use in scientific research and analysis. It serves as a versatile tool for various applications within the laboratory setting. The core function of PIM447 is to facilitate precise measurements and data collection during experimental procedures.

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Lab products found in correlation

Doxycycline, by Merck Group (2 mentions) Dimethyl sulfoxide (dmso), by Thermo Fisher Scientific (2 mentions) Vinorelbine, by Merck Group (1 mentions) H1299 cells, by American Type Culture Collection (1 mentions) Lcl161, by Selleck Chemicals (1 mentions) Paclitaxel, by LC Laboratories (1 mentions) Doxorubicin, by LC Laboratories (1 mentions) Binimetinib, by LC Laboratories (1 mentions) Osi 906, by LC Laboratories (1 mentions) Byl719, by LC Laboratories (1 mentions) Cycloheximide (chx), by Selleck Chemicals (1 mentions) Etomoxir, by Selleck Chemicals (1 mentions) Gw6471, by Selleck Chemicals (1 mentions) Lipidspot 488, by Biotium (1 mentions) Sytox green nucleic acid stain, by Thermo Fisher Scientific (1 mentions) Dialyzed fbs, by Thermo Fisher Scientific (1 mentions) Lipidspot 610, by Biotium (1 mentions) Cycloheximide (chx), by Avantor (1 mentions) P irs1 s1101, by Cell Signaling Technology (1 mentions) Hif1a, by Cell Signaling Technology (1 mentions)

7 protocols using pim447

Combination Therapy for Lung Cancer

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Doxorubicin, OSI-906, BYL719, binimetinib, and paclitaxel were purchased from LC Laboratories (Woburn, MA). PIM447 and LCL161 were obtained from Selleck Chemicals (Houston, TX). Vinorelbine was purchased from Sigma-Aldrich (St. Louis, MO). THZ1 was purchased from Cayman Chemical Company (Ann Arbor, MI). Human lung carcinoma PC9 cells were obtained from Sigma-Aldrich, and H1299 cells were provided from ATCC (Manassas, VA). All cell lines were grown in RMPI-1640 medium supplemented with 10% fetal bovine serum and 1% penicillin–streptomycin, at 37 °C and 5% CO₂.

Bae S.Y., Guan N., Yan R., Warner K., Taylor S.D, & Meyer A.S. (2020). Measurement and models accounting for cell death capture hidden variation in compound response. Cell Death & Disease, 11(4), 255.

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Multiparametric Lipid Imaging Assay

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The following chemicals were used at the indicated concentrations: 20, 50, and 100 ng/ml Doxycycline (Sigma-Aldrich, D5207), 50 nM CHIR (Selleckchem, S2745), 3 µM PIM447 (Selleckchem, S7985), 4 µM GW6471 (Selleckchem, S2798), 100 µM Etomoxir (Selleckchem, S8244), 20 µg/ml cycloheximide (Selleckchem, S7418), 1:1000 LipidSpot488 (Biotium, 70065) or LipidSpot610 (Biotium, 70069), 167 nM SyTOX Green Nucleic Acid Stain (Fisher Scientific, S7020), 25 mM Glucose (Thermo Scientific, A24940-01), 10% Dialyzed FBS (Gibco, A33820-01).

Chauhan S.S., Casillas A.L., Vizzerra A.D., Liou H., Clements A.N., Flores C.E., Prevost C.T., Kashatus D.F., Snider A.J., Snider J.M, & Warfel N.A. (2023). PIM1 drives lipid droplet accumulation to promote proliferation and survival in prostate cancer. Oncogene, 43(6), 406-419.

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Comprehensive Protein Expression Analysis

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Trypsin-EDTA (Cat# 25-053-CI), PBS (Cat# 21-031-CV), and laminin (Cat# 354232) were purchased from Corning. Recombinant EGF was purchased from Life Technologies (Cat# PHG0311). IGF was purchased from Sigma-Aldrich (Cat# I3769). PIM447 was purchased from Selleck Chemicals (Cat# S7985). AZD1208 was acquired from AdooQ Bioscience (Cat# A13203-750). DMSO was purchased from Thermo Fisher Scientific (Cat# 97064-724). Cycloheximide was purchased from VWR (Cat# 97064-724), and doxycycline was purchased from Sigma-Aldrich (Cat# D9891-5G). Recombinant myelin basic protein (MBP) was a gift from Dr. Greg Rogers, and recombinant ABI2 protein was purchased from Origene (Cat# TP300637). Radio-labeled ATP was purchased from Perkin Elmer (Cat# BLU502A). The antibody to ABI2 was purchased from Bethyl Laboratories (Cat# A302-499A-M). GFP (Cat# 2956S), HA (Cat# 3724S), HIF-1a (Cat# 14179S), p-IRS1 [S1101] (Cat# 2385S), PIM1 (Cat# 3247S), and WAVE2 (Cat# 3659S) antibodies were purchased from Cell Signaling Technology. The antibody for actin was purchased from BD Biosciences (Cat# 61656). PIM1 (Cat# ab75776) and Ki67 (Cat#ab833) antibodies used for immunohistochemistry were purchased from Abcam.

Jensen C.C., Clements A.N., Liou H., Ball L.E., Bethard J.R., Langlais P.R., Toth R.K., Chauhan S.S., Casillas A.L., Daulat S.R., Kraft A.S., Cress A.E., Miranti C.K., Mouneimne G., Rogers G.C, & Warfel N.A. (2023). PIM1 phosphorylates ABI2 to enhance actin dynamics and promote tumor invasion. The Journal of Cell Biology, 222(6), e202208136.

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Antibody and Reagent Sources for Cell Signaling

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Rabbit polyclonal anti-CD133 antibody (18470–1-AP) was obtained from Proteintech Group, Inc. (18470–1-AP, Rosemont, IL) and rabbit polyclonal anti-cleaved poly (ADP-ribose) polymerase (PARP) was from EMD Millipore (AB3565, Temecula, CA). Rabbit polyclonal anti-PARP (9542), anti-cleaved caspase 3 (9661), anti-caspase 3 (9662), and rabbit monoclonal anti-vinculin (13901S), anti-β-actin (3700) were from Cell Signaling Technology (Beverly, MA). PIM447 was obtained from Selleckchem (S7985, Houston, TX) and cisplatin was from Cayman Chemical Company (13119, Ann Arbor, MI).

Wadhwani N., Markert H.R., Marayati R., Bownes L.V., Quinn C.H., Aye J.M., Stewart J.E., Yoon K.J, & Beierle E.A. (2021). PIM447 inhibits oncogenesis and potentiates cisplatin effects in hepatoblastoma. Journal of pediatric surgery.

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Evaluation of PIM and FLT3 Inhibitors

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Human PMBL cells Karpas1106P and U2940 and L-428 cHL cells were maintained in RPMI 1640 medium supplemented with 20% fetal bovine serum (Lonza Group AG, Basel, Switzerland). SUP-HD1 cHL cells were grown in McCoy's 5A medium supplemented with 20% fetal bovine serum (Lonza Group AG). B-cell acute lymphoblastic leukemia SEM cell line was cultured in Iscove's modified Dulbecco's medium supplemented with 10% fetal bovine serum (Lonza Group AG). The dual pan-PIM-FMS-like tyrosine kinase 3 (FLT3) inhibitor MEN1703 was provided by Menarini Ricerche (Pomezia, Italy), and the pan-PIM inhibitor PIM447 was purchased from Selleckchem (Houston, TX). All chemicals were dissolved in dimethyl sulfoxide (DMSO) (Sigma-Aldrich, St. Louis, MO). In vehicle-control experiments, final DMSO concentrations was 0.5% (control for a 5-mmol/L dose).

Szydłowski M., Dębek S., Prochorec-Sobieszek M., Szołkowska M., Tomirotti A.M., Juszczyński P, & Szumera-Ciećkiewicz A. (2021). PIM Kinases Promote Survival and Immune Escape in Primary Mediastinal Large B-Cell Lymphoma through Modulation of JAK-STAT and NF-κB Activity. The American journal of pathology, 191(3).

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Cell Viability Assay with Drug Combinations

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Cells were seeded at 2,500 cells per well of 96-well plates 24 hours prior to treatments. To generate GI₅₀ curves, cells were treated with vehicle (DMSO) or serial dilutions of each drug for 72 hours. Each treatment condition was completed in technical triplicate. Cell viability was read-out using CellTiter Glo (Promega G7573) on a BioTek Synergy2 plate reader. The total number of live cells for each drug dose were normalized to the vehicle control and GI₅₀ drug curves were established using GraphPad Prism. For combinations, one drug was added at a constant concentration across all wells and total live cell counts were normalized to the secondary drug-only condition. For three-day growth curves, cells were seeded at 2,500 cells per well of 96-well plates 24 hours prior to treatment and treated with stable concentrations of each drug. Total live cell counts were measured using CellTiter Glo on each day, and relative luciferase units were plotted to represent cell growth over time. Drugs used include BYL719 (Selleckchem S2814) and A-1331852 (Chemietek 1430844–80-6), PIM447 (Selleckchem S7985) and Dasatinib (ApexBio A3017).

Stanland L.J., Ang H.X., Hoj J.P., Chu Y., Tan P., Wood K.C, & Luftig M.A. (2023). CBF-beta mitigates PI3K-alpha-specific inhibitor killing through PIM1 in PIK3CA mutant gastric cancer. Molecular cancer research : MCR, 21(11), 1148-1162.

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Pharmacological Inhibition of PIM Kinases

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Human DLBCL cell lines DHL4, DHL6, U2932, and Burkitt lymphoma (BL) cell line Raji were maintained in RPMI-1640 medium supplemented with 10% FBS, and DLBCL cell lines LY1, LY7, HBL-1, and B-ALL cell line SEM were grown in IMDM supplemented with 10% FBS. The pan-PIM inhibitor MEN1703 was provided by Menarini Ricerche. Pan-PIM inhibitors SGI-1776 and PIM447, MYC inhibitor 10058-F4, and proteasome inhibitor MG132 were purchased from Selleckchem. All chemicals were dissolved in DMSO (Sigma-Aldrich). In vehicle-control experiments, maximal final DMSO concentration was 0.1% (control for 10 mmol/L dose).

Szydłowski M., Garbicz F., Jabłońska E., Górniak P., Komar D., Pyrzyńska B., Bojarczuk K., Prochorec-Sobieszek M., Szumera-Ciećkiewicz A., Rymkiewicz G., Cybulska M., Statkiewicz M., Gajewska M., Mikula M., Gołas A., Domagała J., Winiarska M., Graczyk-Jarzynka A., Białopiotrowicz E., Polak A., Barankiewicz J., Puła B., Pawlak M., Nowis D., Golab J., Tomirotti A.M., Brzózka K., Pacheco-Blanco M., Kupcova K., Green M.R., Havranek O., Chapuy B, & Juszczyński P. (2021). Inhibition of PIM Kinases in DLBCL Targets MYC Transcriptional Program and Augments the Efficacy of Anti-CD20 Antibodies. Cancer research, 81(23).

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