Bay80 6946

Manufactured by Selleck Chemicals

Sourced in United States

BAY80-6946 is a chemical compound used in laboratory research applications. It is a potent and selective inhibitor of the phosphoinositide 3-kinase (PI3K) enzyme. The compound can be utilized by researchers to study the role of PI3K in various biological processes.

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5 protocols using bay80 6946

Quantifying PD-L1 Expression by Flow Cytometry

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Flowcytometric analysis of PD-L1 expression was performed as described previously(25 (link)). Briefly, cells at 70–80% confluency were trypsinized to obtain a single cell suspension. Approximately 500,000 cells were washed with 2% FBS and re-suspended in 100ul 2% FBS and incubated with mouse anti-human PD-L1 (CD274) antibody that was directly conjugated with PE (1:200 dilution, Clone MIH1, BD Pharmingen, San Diego CA) for 30 min at 4°C. Cells were then washed in 2% FBS and analyzed using a Gallios flow cytometer, (Beckman Coulter, Brea CA) and FlowJo software (v.10.1). Prior to collection cells were treated with either Bay 80-6946 (PI3 kinase inhibitor) or SGI-7079 (Axl inhibitor) (both from Selleck Chemical, Houston TX) for 24h where indicated. Immunoblot analysis was performed as described previously(8 ). Commercially available siRNAs specific for either Axl or p110 PI3 kinase (GE Dharmacon, Lafayette CO) were transfected via electroporation (Nucleofector II, Amaxa, Basel, Switzerland) by using program T-001 with transfection agent T (Lonza, Visp, Switzerland). Cells were collected 48–72h post-transfection and assayed as described above. Comparisons between treatment groups were performed using Student’s t-test, with p<0.05 denoting significance.

Skinner H.D., Giri U., Yang L.P., Kumar M., Liu Y., Story M.D., Pickering C.R., Byers L.A., Williams M.D., Wang J., Shen L., Yoo S.Y., Fan Y.H., Molkentine D.P., Beadle B.M., Meyn R.E., Myers J.N, & Heymach J.V. (2017). Integrative analysis identifies a novel AXL-PI3 kinase-PD-L1 signaling axis associated with radiation resistance in head and neck cancer. Clinical cancer research : an official journal of the American Association for Cancer Research, 23(11), 2713-2722.

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Culturing and Characterizing Human Melanoma Cells

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Human melanoma cell lines have been previously described (Satyamoorthy et al., 2003 (link); Iliopoulos et al., 1989 (link)). Cells were cultured in DMEM supplemented with 5% fetal bovine serum and grown at 37°C in 5% CO₂. The consistency of cellular genotypes and cell line identities were confirmed by DNA fingerprinting, using Coriell’s microsatellite kit. PX-866 was provided by Oncothyreon (Seattle, WA, USA); PLX4720 was supplied by Plexxikon/Roche (Berkeley, CA, USA). GDC-0941, BAY80-6946, AZD6482, LY294002 were purchased from Selleckchem (Houston, TX, USA). All compounds were stored at −20°C in DMSO as 10mM stocks.

Shannan B., Chen Q., Watters A., Perego M., Krepler C., Thombre R., Li L., Rajan G., Peterson S., Gimotty P.A., Wilson M., Nathanson K.L., Gangadhar T.C., Schuchter L.M., Weeraratna A.T., Herlyn M, & Vultur A. (2016). Enhancing the evaluation of PI3K inhibitors through 3D melanoma models. Pigment cell & melanoma research, 29(3), 317-328.

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Pharmacological Inhibition of Key Pathways

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Pharmacological inhibitors of MEK1/2 (Trametinib/GSK1120212), PI3K (Copanlisib/BAY 80-6946), and DNMT1 (GSK3685032) were acquired from Selleckchem. KRAS G12C inhibitor (Sotorasib/AMG-510) was purchased from MedChemExpress.

Kostyrko K., Román M., Lee A.G., Simpson D.R., Dinh P.T., Leung S.G., Marini K.D., Kelly M.R., Broyde J., Califano A., Jackson P.K, & Sweet-Cordero E.A. (2023). UHRF1 is a mediator of KRAS driven oncogenesis in lung adenocarcinoma. Nature Communications, 14, 3966.

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In Vivo Evaluation of Targeted Therapies for Prostate Cancer

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All studies performed in mice were approved by the Institutional Animal Care and Use Committee (IACUC) at University of Chicago and performed in compliance with NIH guidelines. Prostate-specific PTEN/p53-deficient (Pb-Cre;PTEN fl/fl Trp53 fl/fl ) mice were screened for autochthonous prostate tumor development at 16 weeks of age by ultrasound. Following the development of solid tumors (5 mm long-axis diameter of solid tumor with ultrasound imaging), the mice were treated with either copanlisib (14mg/kg, iv, every alternate day, Selleckchem; BAY 80-6946), trametinib (3mg/kg, po, every alternate day, Selleckchem; GSK 1120212), LGK`974 (3mg/kg, po, every alternate day, Selleckchem; WNT`974) single agents or their combinations (as indicated in Figure legends). Tumor volumes were monitored using MRI and quantified using Amira software (RRID:SCR_007353), as previously described (24) . If solid tumor volume did not grow >20% (stable disease, SD) at indicated time point following treatment, relative to baseline, mice were considered treatment responsive. Percent Partial response (PR) was calculated based on a proportion of total mice that exhibited >30% decrease in solid tumor volume following treatment, relative to baseline. % Overall Response Rate (ORR) was calculated based on number of responder mice (SD + PR), relative to total mice enrolled in a specific treatment.

Chaudagar K., Hieromnimon H.M., Kelley A., Labadie B., Shafran J., Rameshbabu S., Drovetsky C., Bynoe K., Solanki A., Markiewicz E., Fan X., Loda M, & Patnaik A. (2023). Suppression of Tumor Cell Lactate-generating Signaling Pathways Eradicates Murine PTEN/p53-deficient Aggressive-variant Prostate Cancer via Macrophage Phagocytosis. Clinical cancer research : an official journal of the American Association for Cancer Research, 29(23).

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Synthesis and Characterization of Inhibitors

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Dabrafenib (purity > 99% by HPLC), sorafenib (purity > 99% by HPLC), JNK inhibitor VIII (purity > 99% by HPLC), trametinib (purity > 99% by HPLC), BAY80-6946 (purity > 99% by HPLC), and SCH772984 (purity > 99% by HPLC) were purchased from Selleckchem (Houston, TX, USA). N⁶,2′-O-Dibutyryl adenosine 3’,5′-cyclic monophosphate (purity > 96% by HPLC) and forskolin (purity > 98% by HPLC) were purchased from Merck (Kenilworth, NJ, USA).

Park K., Shin Y., Lee G., Park H, & Choi Y. (2021). Dabrafenib Promotes Schwann Cell Differentiation by Inhibition of the MEK-ERK Pathway. Molecules, 26(8), 2141.

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