Ly294002 ab120243

Manufactured by Abcam

Sourced in United States, United Kingdom

LY294002 (ab120243) is a small molecule that functions as a potent and selective inhibitor of phosphoinositide 3-kinase (PI3K). It is a widely used tool compound for studying the PI3K signaling pathway.

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6 protocols using ly294002 ab120243

Nrf2-Mediated Antioxidant Pathway Regulation

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KGF-2 was provided by the School of Pharmacy, Wenzhou Medical University (Wenzhou, China). Antibodies of p-Akt (#4060), Akt (#4685), cleaved caspase-3 (#9664), Bcl-2 (#4223), and Bax (#2772) were provided by Cell Signaling Technology (Danvers, MA, USA). Antibodies of Nrf2 (ab137550) and the PI3K/Akt inhibitor LY294002 (ab120243) were provided by Abcam (Cambridge, MA, USA). Antibodies of β-actin (20536-1-AP), HO-1 (27282-1-AP), NQO-1 (11451-1-AP), CAT (21260-1-AP), SOD2 (24127-1-AP), and horseradish peroxidase-conjugated goat antimouse/rabbit secondary antibodies (SA00001-1 or SA00001-2) came from ProteinTech (Rosemont, USA). Hydrogen peroxide (H₂O₂, #349887), zinc protoporphyrin IX (ZnPP, #282820) and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT, M2128), and dimethyl sulfoxide (DMSO, D8418) came from Sigma-Aldrich (St. Louis, MO, USA). The reagents of cell culture were obtained from Gibco (Grand Island, NY, USA). The fluorescent dyes 2′,7′-dichlorodihydrofluorescein diacetate (DCFH-DA, S0033) and Annexin V-FITC/PI apoptosis detection kit (C1062L) were provided by Beyotime (Shanghai, China). Nrf2-small interfering RNA and fluorescein-conjugated control siRNA (sc-37030 and sc-36869) were provided by Santa Cruz Biotechnology Inc. (CA, USA).

Liu S., Jin Z., Xia R., Zheng Z., Zha Y., Wang Q., Wan X., Yang H, & Cai J. (2022). Protection of Human Lens Epithelial Cells from Oxidative Stress Damage and Cell Apoptosis by KGF-2 through the Akt/Nrf2/HO-1 Pathway. Oxidative Medicine and Cellular Longevity, 2022, 6933812.

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Quantitative Western Blot Analysis

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Proteins were extracted from frozen heart tissues. The bicinchoninic acid protein assay was used to determine the protein concentration using bovine serum albumin as a standard (Pierce Rockford, IL). Equal amounts of proteins were loaded on SDS-PAGE gels; after separation, the proteins were transferred to a polyvinylidene difluoride membrane. The membranes were incubated with anti-MAPK p38 (CST, #8690S), p-MAPK p38 (CST, #4511S), rat anti-AKT (ab8805, Abcam), p-AKT (ab81283, Abcam), rabbit anti-Nrf2 (1:1000, 16396-1-AP, Proteintech), rabbit anti-PCNA (1:1000, 10205-2-AP, Proteintech), rabbit anti-HO-1 (1:1000, #82206S, CST), and rabbit antibody to β-Actin (1:2500, #8457, CST) antibodies. The bands were visualized by enhanced chemiluminescence and scanned using FLUORCHEM™ E (ProteinSimple, United States). The densitometric results were quantified using ImageJ software. The results from western blotting were normalized to the control group.
MAPK inhibitor (SB203580) was purchased from Merck-Millipore (Billerica, MA, United States), and PI3K inhibitor (LY294002, ab120243) was purchased from Abcam.

Chen L., Guo Y., Qu S., Li K., Yang T., Yang Y., Zheng Z., Liu H., Wang X., Deng S., Zhang Y., Zhu X, & Li Y. (2021). The Protective Effects of Shengmai Formula Against Myocardial Injury Induced by Ultrafine Particulate Matter Exposure and Myocardial Ischemia are Mediated by the PI3K/AKT/p38 MAPK/Nrf2 Pathway. Frontiers in Pharmacology, 12, 619311.

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Sulforaphane-Mediated Regulation of NRF2 Pathway

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Sulforaphane (S6317), as well as anti-mouse IgG (HRP) (1:3000, A-9044) and anti-rabbit IgG (HRP) (1:3000, A-0545) antibodies were purchased from Sigma-Aldrich. LY294002 (ab120243) was purchased from Abcam. The rabbit normal IgG and antibodies against NRF2 (1:1000, sc-13032), sMAF (1:1000, sc-22831), NQO1 (1:1000, sc-32793), FN1 (1:1000, sc-8422), COL1A1 (1:1000, sc-293,182), α-SMA (1:1000, sc-53142), N-Cadherin (1:500, sc-7939) and GAPDH (1:3000, sc-32233) were purchased from Santa Cruz Biotechnology. The antibody against E-Cadherin (1:1000, 610,181) was purchased from BD Biosciences. Antibodies against AKT (1:2000, 4691S) and phospho-AKT (Ser473) (1:1000, 4060S) were purchased from Cell Signaling Technology. TGF-β (100–21) was purchased from PeproTech.

Liu P., Luo G., Dodson M., Schmidlin C.J., Wei Y., Kerimoglu B., Ooi A., Chapman E., Garcia J.G, & Zhang D.D. (2020). The NRF2-LOC344887 signaling axis suppresses pulmonary fibrosis. Redox Biology, 38, 101766.

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Investigating Molecular Mechanisms in Cancer

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Antibodies against SNCG (ab55424), Phalloidin (ab176753), Ki-67 (ab15580), F-actin (ab112124) were obtained from Abcam (Cambridge, UK). Antibodies against mTOR (sc-517,464), p-mTOR (Sec2448, sc-293,133), p70S6 kinase (sc-8418), GAPDH (sc-47,724), β-Actin (sc-47,778), Cyclin D1 (sc-8396), MMP9 (sc-393,859, sc-21,733), Vimentin (sc-6260) were purchased from Santa Cruz (CA, USA). Antibodies against p-p70S6 kinase (Thr389, #97596), p-Akt (Sec473, #4060), Akt (#4961), MMP2 (#40994), E-cadherin (#14472), N-cadherin (#13116) were purchased from Cell Signaling Technology. Secondary antibodies, including HRP-labeled goat anti-mouse IgG (A0216), HRP-labeled goat anti-rabbit IgG (A0208), Cy3-labeled goat anti-rabbit IgG (P0183), were purchased from Beyotime (Shanghai, China). LY294002 (ab120243) was obtained from Abcam (Cambridge, UK), Puromycin (sc-205,821) was purchased from Santa Cruz (CA, USA). insulin-like growth factor 1 (IGF-1) (P5502) was purchased from Beyotime (Shanghai, China). DMSO (PYG0040) and DAPI (AR1177) were purchased from Bosterbio (Hubei, China).

Zhang J., Liu X.H., Li C., Wu X.X., Chen Y.L., Li W.W., Li X., Gong F., Tang Q, & Jiang D. (2020). SNCG promotes the progression and metastasis of high-grade serous ovarian cancer via targeting the PI3K/AKT signaling pathway. Journal of Experimental & Clinical Cancer Research : CR, 39, 79.

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Mast Cell Activation Assay

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Dinitrophenyl-human serum albumin (DNP-HSA), anti-DNP-IgE, o-phthaldialdehyde, and 4-nitrophenyl N-acetyl-β-d-glucosamide were purchased from Sigma-Aldrich (St. Louis, MO, USA). The protease inhibitor cocktail and phosphatase inhibitor cocktail were purchased from Roche (Mannheim, Germany). IL-3 and stem cell factor (SCF) were purchased from PeproTech (EC, London, UK). The signaling inhibitors, PP2 and Akt inhibitor IV were purchased from Sigma-Aldrich, and U 73122 and LY 294002 (ab120243) were from Abcam (Cambridge, UK). The antibodies for anti-IRF3 and p-S396-IRF3 were obtained from Cell Signaling (Beverly, MA, USA), lamin B1 was obtained from Santa-Cruz Biotechnology (Dallas, TX, USA), and anti-HDC was purchased from Invitrogen (Waltham, MA, USA).

Choi Y.A., Dhakal H., Lee S., Kim N., Lee B., Kwon T.K., Khang D, & Kim S.H. (2023). IRF3 Activation in Mast Cells Promotes FcεRI-Mediated Allergic Inflammation. Cells, 12(11), 1493.

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Evaluating Vincristine Resistance Mechanisms

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Trypsin (T4549), streptomycin (85886), penicillin (V900929), and vincristine (VCR) were obtained from Sigma-Aldrich (Shanghai, China). MKN-28/VCR cells were cultured in RPMI-1640 medium (Invitrogen, Thermo Fisher Scientific, Waltham, MA, USA) with fetal bovine serum (B8687; Sigma-Aldrich,). Siva-1 (#12532), Poly(C)-binding protein 1 (PCBP1; #8534), Akt (#9272S), NF-κB (#8242), MRP1 (#72202), MDR1 (#13342), and GAPDH (#5174) antibodies were purchased from Cell Signaling Technology, Inc., USA. MTT assay kits (ab211091) and Akt pathway specific inhibitor LY 294002 (ab120243) were purchased from ABCAM (Shanghai, China). Reverse transcriptase-polymerase chain reaction (RT-PCR) kits were purchased from Thermo Fisher Scientific.

KONG F., WU K., PANG L., HUANG Y., LI L., XU J., LI F., QING Y., WANG Z., HUANG X., XU S., ZHONG X., ZHU Z., WANG X, & YANG J. (2023). Inhibition of apoptosis-regulatory protein Siva-1 reverses multidrug resistance in gastric cancer by targeting PCBP1. Oncology Research, 30(6), 277-288.

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