Gsk3β 22104 1 ap

Manufactured by Proteintech

Sourced in China

GSK3β (22104-1-AP) is a primary antibody that recognizes the glycogen synthase kinase 3 beta (GSK3β) protein. GSK3β is a serine/threonine protein kinase involved in the regulation of various cellular processes.

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Lab products found in correlation

Gapdh antibody, by Santa Cruz Biotechnology (1 mentions) Ecl western blot kit, by Cytiva (1 mentions) Cleaved caspase 3 ab2302, by Abcam (1 mentions) β catenin 51067 2 ap, by Proteintech (1 mentions) Cyclind1 60 186 1 ig, by Proteintech (1 mentions) β actin, by Proteintech (1 mentions) Normal igg peroxidase conjugated affinipure goat anti rabbit mouse igg h l, by Jackson ImmunoResearch (1 mentions) Gst 10000 0 ap, by Proteintech (1 mentions) Ha no 3724, by Cell Signaling Technology (1 mentions) β actin 60008 1 lg, by Proteintech (1 mentions) Mg132, by Selleck Chemicals (1 mentions) Tubulin 11224 1 ap, by Proteintech (1 mentions) Gapdh ap0063, by Bioworld Technology (1 mentions) Ab128873, by Abcam (1 mentions) Ab180967, by Abcam (1 mentions) Ki67 ab16667, by Abcam (1 mentions) Pierce co immunoprecipitation kit, by Thermo Fisher Scientific (1 mentions) Ab182141, by Abcam (1 mentions) Ripa lysis buffer, by Beyotime (1 mentions) Phosphatase inhibitor, by Beyotime (1 mentions)

Spelling variants of this product

GSK3β (36 citations)

7 protocols using gsk3β 22104 1 ap

Western Blot Analysis of Protein Markers

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20 μg protein extract were separated on 10% SDS-PAGE and transferred to polyvinylidene difluoride membranes. The membranes were blocked with 5% skimmed milk and followed by incubating with the primary antibody (FUT4, AP12067b, Abgent; cleaved caspase-3 ab2302, Abcam; cleaved PARP, ab4830, Abcam; Sp1, ab13370, Abcam; CD44, ab157107, Abcam; GSK-3β, 22,104–1-AP, Proteintech; p-GSK-3β, 22,104–1-AP, Proteintech; β-catenin, 51,067–2-AP, Proteintech; CyclinD1, 60,186–1-Ig, Proteintech; GAPDH, AP7873a, Abgent) on a shaker overnight at 4 °C. The membranes were then incubated with horseradish peroxidase-conjugated secondary antibody (rabbit IgG, 1/1000 diluted; UK). A GAPDH antibody (1/200 diluted; Santa Cruz Biotech) was used as a control. All bands were detected using ECL Western blot kit (Amersham Biosciences, UK). The bands were measured with LabWorks (TM ver4.6, UVP, BioImaging systems).

Liu B., Ma H., Liu Q., Xiao Y., Pan S., Zhou H, & Jia L. (2019). MiR-29b/Sp1/FUT4 axis modulates the malignancy of leukemia stem cells by regulating fucosylation via Wnt/β-catenin pathway in acute myeloid leukemia. Journal of Experimental & Clinical Cancer Research : CR, 38, 200.

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Quantitative Protein and Gene Expression

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Western blotting and reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) were used for molecules detection and performed as previously described (Wu et al. 2015 (link)). Antibodies against HK2 (22029-1-AP,), AKT1 (10176-2-AP), LDHA (19987-1-AP, ), GLUT1 (21829-1-AP), and GSK3β (22104-1-AP) all purchased from Proteintech; β-Actin (#4970), phosphor-AKT1 (Ser473, #4060), and phosphor-GSK3β (Ser9, #5558), all purchased from Cell Signaling Technology, were used to assess the expression of an individual protein. Secondary antibodies were used at 1:5000 dilutions. The primer sequences for RT-qPCR involved in this study were shown in Supplemental Table 2.

Wang Z., Yang Y., Hu S., He J., Wu Z., Qi Z., Huang M., Liu R., Lin Y., Tan C., Xu M, & Zhang Z. (2020). Short-form RON (sf-RON) enhances glucose metabolism to promote cell proliferation via activating β-catenin/SIX1 signaling pathway in gastric cancer. Cell Biology and Toxicology, 37(1), 35-49.

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Immunoblotting Assay with Proteasome and Protein Synthesis Inhibitors

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The proteasome inhibitor MG132 and protein synthesis inhibitor cycloheximide (CHX) were purchased from Selleck. Antibodies used in immunoblotting: eEF2K (ab45168, ab85721, 1:1000) was purchased from Abcam; PD-L1 (17952-1-AP, 1:1000), GSK3β (22104-1-AP, 1:2000), β-actin (60008-1-lg, 1:5000), Flag (66008-3-lg, 1:5000), tubulin (11224-1-AP, 1:5000), GST (10000-0-AP, 1:4000) were purchased from Proteintech; HA (No. 3724, 1:1000), p-GSK3β/S9 (No. 9323, 1:1000) were purchased from Cell Signaling Technologies; phosphoserine monoclonal antibody (mAb) (CSB-MA080235, 1:1000) was purchased from CUSABIO. Normal IgG/Peroxidase-conjugated AffiniPure Goat Anti-Rabbit/Mouse IgG (H+L) was purchased from Jackson Immuno Research.

Chen X., Wang K., Jiang S., Sun H., Che X., Zhang M., He J., Wen Y., Liao M., Li X., Zhou X., Song J., Ren X., Yi W., Yang J., Chen X., Yin M, & Cheng Y. (2022). eEF2K promotes PD-L1 stabilization through inactivating GSK3β in melanoma. Journal for Immunotherapy of Cancer, 10(3), e004026.

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Antibody Sourcing for EMT Markers

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Antibodies against EFNA4 (19685-1-AP), E-cadherin (60335-1-Ig), N-cadherin (66219-1-Ig), vimentin (10366-1-AP), EPHA2 (66736-1-Ig), and GSK3β (22104-1-AP) were purchased from Proteintech (Wuhan, China). Antibodies against β-catenin (#8480), AKT (#4691), phospho-AKT (Ser473; #4060), phospho-GSK3β (Ser9; #9323), and EPHA2 (#6997) were obtained from Cell Signaling Technology (Beverly, MA, USA). Antibodies against Ki67 (ab16667), PIK3R2 (ab180967), and CTCF (ab128873) were obtained from Abcam (Cambridge, MA, USA). The antibody against glyceraldehyde 3-phosphate dehydrogenase (GAPDH; AP0063) was purchased from Bioworld Technology (Bloomington, MN, USA). Human EFNA4 antibody (MAB3692) was purchased from Bio-Techne (Minneapolis, MN, USA). Phospho-EPHA2 (Ser897; AP1082) and phospho-EPHA2 (Tyr772; AP0817) were purchased from ABclonal Technology (Wuhan, China). Monoclonal anti-FLAG M2 antibody (1804) was obtained from Merck KGaA (Darmstadt, Germany).

Lin J., Zeng C., Zhang J., Song Z., Qi N., Liu X., Zhang Z., Li A, & Chen F. (2021). EFNA4 promotes cell proliferation and tumor metastasis in hepatocellular carcinoma through a PIK3R2/GSK3β/β-catenin positive feedback loop. Molecular Therapy. Nucleic Acids, 25, 328-341.

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Protein Extraction and Purification

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Protein extraction and purification were performed using Pierce™ Co-Immunoprecipitation Kit (26149, Thermo Fisher Scientific). Mass spectrometry was performed by Novogene (Beijing, China) and results were provided in Supplementary Table 3. The primary antibodies included PTGDS (ab182141, Abcam), MYH9 (60233-1-Ig, Proteintech Group), and GSK3-β (22104-1-AP, Proteintech Group).

Hu S., Ren S., Cai Y., Liu J., Han Y., Zhao Y., Yang J., Zhou X, & Wang X. (2021). Glycoprotein PTGDS promotes tumorigenesis of diffuse large B-cell lymphoma by MYH9-mediated regulation of Wnt–β-catenin–STAT3 signaling. Cell Death and Differentiation, 29(3), 642-656.

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Astragaloside A, Chlorogenic Acid, and Scutellarin Protocol

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Astragaloside A, sourced from Shanxi Qingya Biological (Co., Ltd., AIV-98-210902, purity≥ 98%), Chlorogenic acid from Xian Xinlu Biotechnology (Co., Ltd., EK-CHA2021091801, purity≥ 98%), and Scutellarin from Kangzhou Biotechnology (Co., Ltd., KZ-2021041201, purity≥ 98%). Naoluotong (NLT) Capsule was obtained from Guangdong Yunfang Pharmaceutical with national medicine permission number Z20063147 and batch number 20210601. Chloral hydrate was supplied by Chengdu Kelong Chemical Reagent Factory with batch number 190420. Benzylpenicillin Potassium for Injection was purchased from Jiangxi Keda Animal Pharmaceutical (Co., Ltd., 20190102).
PMSF, Phosphatase inhibitor, BCA Protein Assay Kit, RIPA lysis buffer, LY294002 (inhibitor of phosphatidylinositol 3-kinase, S1737-25 mg) and cleaved Caspase 3 rabbit monoclonal antibody (AF1150, 1: 1000) were provided by Beyotime Biotechnology (Shanghai, China). SOD, MDA, GSH-Px, CAT, ChE, CHAc assay (ELISA) kits were supplied by Enzyme-Linked Biotechnology (Shanghai). PI3K (20584-1-AP, 1: 500), AKT (60203-2-Ig, 1: 5000), p-AKT (66444-1-Ig, 1: 5000), GSK3β (22104-1-AP, 1: 4000), Bax (60267-1-Ig, 1: 10000), Bcl-2 (26593-1-AP, 1: 2000) and Cyto-c (10993-1-AP, 1: 4000) were supplied by Proteintech (Wuhan, China); p-PI3K (AF3242, diluted 1: 500) and p-GSK3β (AF2016, diluted 1: 1000) were provided by Affinity Biosciences (Cincinnati, OH, USA).

Cheng F., Zhang J., Yang P., Chen Z., Fu Y., Mi J., Xie X., Liu S, & Sheng Y. (2024). Exploring the neuroprotection of the combination of astragaloside A, chlorogenic acid and scutellarin in treating chronic cerebral ischemia via network analysis and experimental validation. Heliyon, 10(8), e29162.

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Comprehensive Protein Analysis Protocol

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Total protein extraction and western blotting were performed following standard methods [15 (link)]. The nuclear and cytoplasmic proteins were extracted with NE-PER nuclear and cytoplasmic extraction reagents (Thermo Fisher Scientific, MA, USA). The primary antibodies included PTGDS (ab182141, Abcam; sc-390717, Santa Cruz Biotechnology), MYH9 (11128-1-AP, 60233-1-Ig, Proteintech Group), TRAF6 (66498-1-Ig, Proteintech Group), GSK3-β (22104-1-AP, Proteintech Group), and other antibodies bought from Cell Signaling Technology (Beverly, USA), including c-myc (18583), Cyclin D1 (2922), CDK2 (2546), caspase 3 (9662), caspase 9 (9508), PARP (9532), Bax (5023), Bcl-xl (2764), zeb-1(3396), vimentin (5741), P21 (2947), p-H2AX (Ser139, 9718), LRP6 (2560), p-LRP6 (Ser1490, 2568), p-GSK3-β (Ser9, 5558), β-catenin (8480), TCF4 (2569), p-STAT3 (Tyr705, 9145), STAT3 (9139), Ubiquitin (3936), His (12698), Flag (14793), and HA (3724). β-tubulin (86298), Histone H3 (4499), and GAPDH (97166) were served as the internal reference.

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