Gsk3β

Manufactured by Bioss Antibodies

Sourced in China

GSK3β is a serine/threonine protein kinase that plays a key role in the regulation of various cellular processes, including glycogen metabolism, gene transcription, and cellular signaling pathways. It is involved in the Wnt/β-catenin signaling pathway and is a critical regulator of cellular homeostasis.

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Lab products found in correlation

β catenin, by Bioss Antibodies (2 mentions) Phospho gsk3β, by Bioss Antibodies (2 mentions) Pvdf membrane, by Merck Group (1 mentions) Horseradish peroxidase labeled secondary antibody, by Abcam (1 mentions) β actin, by Abcam (1 mentions) Enhanced chemiluminescence reagent, by Thermo Fisher Scientific (1 mentions) Vimentin, by Bioss Antibodies (1 mentions) Chemidoc crs molecular imager, by Bio-Rad (1 mentions) Bca protein assay kit, by Thermo Fisher Scientific (1 mentions) Cyclin d1, by Bioss Antibodies (1 mentions) P akt, by Bioss Antibodies (1 mentions) C jun, by Bioss Antibodies (1 mentions) Hmga1, by Cell Signaling Technology (1 mentions) P pi3k, by Cell Signaling Technology (1 mentions) Cyclin d1, by Cell Signaling Technology (1 mentions) E cadherin, by Cell Signaling Technology (1 mentions) β actin, by Proteintech (1 mentions) β catenin, by Cell Signaling Technology (1 mentions) P gsk3β ser9, by Abcam (1 mentions) Vimentin, by Cell Signaling Technology (1 mentions)

Spelling variants of this product

Anti-GSK3β (2 citations) Phospho-GSK3β (2 citations)

4 protocols using gsk3β

Protein Extraction and Western Blot Analysis

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Total protein was isolated from BMECs using a tissue protein extraction kit (Nanjing KeyGEN Biotech. Co. Ltd., Nanjing, China), and protein concentration was measured using a BCA assay kit (Nanjing KeyGEN). The protein loading volume was adjusted to 30 μg/well. Proteins were subjected to gel electrophoresis (SDS-PAGE, 12.5%) and transferred to polyvinylidene fluoride (PVDF) membranes (Sigma-Aldrich). PVDF membranes were incubated with β-catenin, GSK3β, phospho-GSK3β (Bioss, Beijing, China), axin-1, TCF-4, cyclinD1 (Biosciences, Shanghai, China), and β-actin (Abcam, Cambridge, UK) antibodies for 12 h at 4°C and washed 3 times with TBST, before incubation with horseradish peroxidase-labeled secondary antibodies (Abcam) for 2 h at 25°C and a final thrice wash with TBST. Exposure and capturing of images were performed using an ECL chemiluminescent color development solution (Shanghai Epizyme Biomedical Technology Co., Ltd, Shanghai). Grayscale values were analyzed using the ImageJ software.

Fan J., Jia F., Liu Y, & Zhou X. (2022). Astragalus polysaccharides and astragaloside IV alleviate inflammation in bovine mammary epithelial cells by regulating Wnt/β-catenin signaling pathway. PLoS ONE, 17(7), e0271598.

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Western Blot Protein Analysis

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Cell-lysate protein concentrations were determined using a BCA protein assay kit (Thermo Scientific, Waltham, MA, USA). Proteins were separated by SDS-PAGE and transferred onto polyvinylidene fluoride membranes, which were probed with primary antibodies: mouse polyclonal Akt, GSK-3β, CDK4, and β-catenin (1:1000; Bioss, Beijing, China); rabbit polyclonal p-Akt, vimentin, cyclin D1, MYH9, c-Jun, vimentin, Sox2, and Oct-4 (1:1000; Bioss, Beijing, China), and rabbit polyclonal PI3K, p-PI3K, and HMGA1 (1:1000; Cell Signaling Technology, Danvers, USA). Rabbit monoclonal anti-β-actin antibody (1:1000; CoWin Bioscience, Beijing, China) was used for normalization. HRP-conjugated anti-rabbit or anti-mouse IgG antibodies were used as secondary antibodies (1:2000; CoWin Bioscience, Beijing, China). Proteins were detected using an enhanced chemiluminescence reagent (Thermo Scientific, Waltham, MA, USA). Images were captured using a ChemiDoc^TM CRS + Molecular Imager (Bio-Rad, Hercules, CA, USA). Grayscale semi-quantification of the bands is shown in Supplemental Fig. 4 for all western blots.

Que T., Zheng H., Zeng Y., Liu X., Qi G., La Q., Liang T., Li Z., Yi G., Zhang S., Li J., Nie J., Tan J.E, & Huang G. (2021). HMGA1 stimulates MYH9-dependent ubiquitination of GSK-3β via PI3K/Akt/c-Jun signaling to promote malignant progression and chemoresistance in gliomas. Cell Death & Disease, 12(12), 1147.

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Comprehensive Immunoblotting Assay Protocol

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The primary antibodies recognized as following: CDK6 (Proteintech, 19117-1-AP), p27 (Proteintech, 25614-1-AP), MMP2 (Proteintech, 10373-2-AP), MMP9 (Proteintech, 10375-2-AP), Histone H3 (Proteintech, 17168-1-AP), β-actin (Proteintech, 60008-1-Ig), GSK3β (Bioss, bs-0023M), p-GSK3β (Ser9) (abcam, ab75745), cyclin D1 (Cell Signaling Technology, #2922), pro-caspase-3/cleaved caspase-3 (Cell Signaling Technology, #9662), pro-PARP/cleaved PARP (Cell Signaling Technology, #9532), E-cadherin (Cell Signaling Technology, #14472), Vimentin (Cell Signaling Technology, #5741), β-catenin (Cell Signaling Technology, #8480), Ki-67 (Abclonal, A2094). The secondary antibodies were purchased from Proteintech (Rosemont, IL, USA). OMT was purchased from Aladdin regents (A111285). Taxol was obtained from Aladdin regents (P106869).

Jin Y., Liu J., Liu Y., Liu Y., Guo G., Yu S, & An R. (2020). Oxymatrine Inhibits Renal Cell Carcinoma Progression by Suppressing β-Catenin Expression. Frontiers in Pharmacology, 11, 808.

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Moxifloxacin-Induced Apoptosis and Autophagy

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MOX was purchased from Sigma-Aldrich/Merck KGaA (Darmstadt, Germany; European Pharmacopoeia Reference Standard), and a purity of > 95%. MOX was dissolved in dimethyl sulfoxide (DMSO). FBS was provided by Zhejiang Dayhang biological technology (Hangzhou, China). Penicillin-streptomycin Solution, Annexin V-FITC Apoptosis Detection Kit was purchased from Beyotime Biotechnology (Shanghai, China). MTT, diaminobenzidine (DAB), polyvinylidene fluoride (PVDF) membranes and hematoxylin were purchased from Sigma-Aldrich/Merck KGaA. The reagents 3-methyladenine (3-MA), chloroquine (CQ) and DMSO were obtained from Sigma-Aldrich/Merck KGaA. Antibodies used in this study were against LC3B (CST, 2775S, USA), P62 (CST, 5114T, USA), AKT (CST, 9272S, USA), Phospho-AKT (Ser473) (Beyotime, AA329, China), mTOR (CST, 2972S, USA), Phospho-mTOR (S2448) (Boster, BM4840, China), p70S6K (Bioss, bs-6370R, China), Phospho-p70S6K (Ser417) (Bioss, bs-5668R, China), GSK3β (Bioss, bs-0023RR, China), Phospho-GSK3β (Bioss, bs-0028RR, China), Ki67 (Bioss, bs-23103, China), β-actin (Sigma-Aldrich, A1978, Germany), goat anti-rabbit immunoglobulin G (IgG) (H+L)-horseradish peroxidase (HRP; SunGene GmbH, LK2001, Germany) and goat anti-mouse IgG (H+L)-HRP (OriGene Technologies, ZB-2305, USA).

Liu J., Liang H., Khilji S., Li H., Song D., Chen C., Wang X., Zhang Y., Zhao N., Li X, & Gao A. (2020). Moxidectin induces Cytostatic Autophagic Cell Death of Glioma Cells through inhibiting the AKT/mTOR Signalling Pathway. Journal of Cancer, 11(19), 5802-5811.

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