GSK-3 alpha/beta [p Ser9, p Ser21, p Ser9] Antibody [Unconjugated]

Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line untreated (-) or treated (+) with 200 nM PMA for 20 minutes and MCF-7 human breast cancer cell line untreated or treated with 100 ng/mL Recombinant Human IGF-1 (291-G1) for 15 minutes. PVDF membrane was probed with 0.2 µg/mL of Human/Mouse/Rat Phospho-GSK-3a/ beta (S21/S9) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1590), followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (HAF008). Specific bands were detected for Phospho-GSK-3a (S21) and GSK-3 beta (S9) at approximately 51 and 46 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Phospho-GSK‑3 alpha / beta (S21/S9) was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line treated with PMA (left panel; positive staining) and untreated HeLa cell line (right panel; negative control) using Rabbit Anti-Human/Mouse/Rat Phospho-GSK‑3 alpha / beta (S21/S9) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1590) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; NL004) and counterstained with DAPI (blue). Specific staining was localized to nuclei. Staining was performed using our protocol for Fluorescent ICC Staining of Non-adherent Cells.

HYS-32 modulates GSK3 beta phosphorylation.(A) Cell lysates from control astrocytes (Con) or astrocytes treated for 2, 6, 12, or 24 h with 5 μM HYS-32 were subjected to 10% SDS-PAGE, and analyzed by immunoblotting with antibodies against GSK3 beta -pTyr216, GSK3 beta -pSer9, or GAPDH. (B) Densitometric analyses of GSK3 beta -pTyr216 and GSK3 beta -pSer9 expressed as the density of the bands in the treated groups relative to the control. *p<0.05 compared to control using one-way ANOVA with Dunnett’s post-hoc test. The results were collected from five independent experiments. (C) According to the data in (B), the stacked bar graph showing the relative percentage of phospho-GSK3 beta at indicated time. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0126217), licensed under a CC-BY license. Not internally tested by R&D Systems.

LY294002 inhibits the HYS-32-induced phosphorylation of GSK3 beta -pS9 and GSK3 beta -pY216.(A) Control astrocytes (Con) or astrocytes treated for 24 h with 5 μM HYS-32 (HYS), co-treated for 24 h with 5 μM HYS-32 and 20 μM LY294002 (HYS+LY), or treated with 20 μM LY294002 (LY) were subjected to 10% SDS-PAGE, and analyzed by immunoblotting with antibodies against GSK3 beta -pSer9, GSK3 beta -pY216, totalGSK3 beta , or GAPDH. (B) Densitometric analyses of GSK3 beta -pSer9 and GSK3 beta -pY216 expressed as the density of the bands in the treated groups relative to the control. (C) According to the data in (B), the stacked bar graph showing relative percentage ofphospho-GSK3 beta in various treatment. (D)Astrocytes treated as in (A) were fixed in cold acetone and triple-stained for beta -tubulin, N-cadherin, and F-actin. Quantitative analysis of the straight distance between microtubule tips and cell border were performed as described in Materials and Methods. The results were collected from three independent experiments.*p<0.01 compared to HYS using one-way ANOVA with Dunnett’s post-hoctest. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0126217), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Hu, Mu, Rt
• Applications: WB, ICC/IF
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: LYOPH

GSK-3 alpha/beta [p Ser9, p Ser21, p Ser9] Antibody [Unconjugated] Summary

• Immunogen: Phosphopeptide containing GSK-3 beta S9 site
• Modification: p Ser9, p Ser9, p Ser21
• Specificity: Detects human, mouse, and rat GSK-3 beta when phosphorylated at S9, and human, mouse, and rat GSK‑3 alpha when phosphorylated at S21.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: GSK3A
• Purity Statement: Antigen and protein A Affinity-purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 5-15 ug/mL
  • Western Blot 0.2 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details.
  *Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for GSK-3 alpha/beta [p Ser9, p Ser21, p Ser9] Antibody [Unconjugated]

• DKFZp686D0638
• EC 2.7.11
• EC 2.7.11.26
• glycogen synthase kinase 3 alpha
• glycogen synthase kinase-3 alpha
• GSK-3 alpha
• GSK-3 alpha/beta

Background

GSK-3 is a Ser/Thr kinase first identified as an inactivator of Glycogen Synthase. GSK-3 acts as a multifunctional downstream switch that determines the output of numerous signaling pathways. There are two mammalian GSK-3 isoforms encoded by distinct genes, GSK-3 alpha and GSK-3 beta, which are structurally similar, but functionally non-identical. GSK-3a is inhibited by phosphorylation at S21 by Akt and other kinases. GSK-3 alpha and GSK-3 beta share 85% amino acid identity. Dysregulated GSK-3 has been implicated in several diseases including type II diabetes, Alzheimer's disease, bipolar disorder, and cancer.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for GSK-3 alpha/beta Antibody (AF1590)(12)

Publications using AF1590

• Shu, Y;Tan, Z;Pan, Z;Chen, Y;Wang, J;He, J;Wang, J;Wang, Y; Inhibition of inflammatory osteoclasts accelerates callus remodeling in osteoporotic fractures by enhancing CGRP+TrkA+ signaling Cell death and differentiation 2024-09-02 [PMID: 39223264] (Western Blot, Transgenic Mouse)
• Kennedy, KE;Kerlero de Rosbo, N;Uccelli, A;Cellerino, M;Ivaldi, F;Contini, P;De Palma, R;Harbo, HF;Berge, T;Bos, SD;Høgestøl, EA;Brune-Ingebretsen, S;de Rodez Benavent, SA;Paul, F;Brandt, AU;Bäcker-Koduah, P;Behrens, J;Kuchling, J;Asseyer, S;Scheel, M;Chien, C;Zimmermann, H;Motamedi, S;Kauer-Bonin, J;Saez-Rodriguez, J;Rinas, M;Alexopoulos, LG;Andorra, M;Llufriu, S;Saiz, A;Blanco, Y;Martinez-Heras, E;Solana, E;Pulido-Valdeolivas, I;Martinez-Lapiscina, EH;Garcia-Ojalvo, J;Villoslada, P; Multiscale networks in multiple sclerosis PLoS computational biology 2024-02-01 [PMID: 38329927] (Flow Cytometry, Human)
• Meah, A;Vedarethinam, V;Bronstein, R;Gujarati, N;Jain, T;Mallipattu, SK;Li, Y;Wang, J; Single-Cell Spatial MIST for Versatile, Scalable Detection of Protein Markers Biosensors 2023-08-27 [PMID: 37754086] (IHC, Mouse)
• S Li, DM Pritchard, LG Yu Galectin-3 promotes secretion of proteases that decrease epithelium integrity in human colon cancer cells Cell Death & Disease, 2023-04-13;14(4):268. 2023-04-13 [PMID: 37055381] (Western Blot, Human)
• Y Tang, KT Kline, XS Zhong, Y Xiao, H Lian, J Peng, X Liu, DW Powell, G Tang, Q Li Chronic colitis upregulates microRNAs suppressing brain-derived neurotrophic factor in the adult heart PLoS ONE, 2021-09-20;16(9):e0257280. 2021-09-20 [PMID: 34543287] (Western Blot, Rat)
• Naphannop Sereesongsaeng, Sara H. McDowell, James F. Burrows, Christopher J. Scott, Roberta E. Burden Cathepsin V suppresses GATA3 protein expression in luminal A breast cancer Breast Cancer Research 2020-12-09 [PMID: 33298139]
• Lan, SY;Tan, MA;Yang, SH;Cai, JZ;Chen, B;Li, PW;Fan, DM;Liu, FB;Yu, T;Chen, QK; Musashi 1-positive cells derived from mouse embryonic stem cells treated with LY294002 are prone to differentiate into intestinal epithelial-like tissues Int. J. Mol. Med. 6/1/2019 [PMID: 30942388]
• Balint Csoboz, Imre Gombos, Eniko Tatrai, Jozsef Tovari, Anna L. Kiss, Ibolya Horvath et al. Chemotherapy induced PRL3 expression promotes cancer growth via plasma membrane remodeling and specific alterations of caveolae-associated signaling Cell Communication and Signaling 2018-12-01 [PMID: 30157875]
• Chiu C, Liao C, Shen C, Tang T, Jow G, Wang H, Wu J HYS-32-Induced Microtubule Catastrophes in Rat Astrocytes Involves the PI3K-GSK3beta Signaling Pathway. PLoS ONE, 2015-05-04;10(5):e0126217. 2015-05-04 [PMID: 25938237] (Western Blot, Rat)
• Lutay N, Hakansson G, Alaridah N, Hallgren O, Westergren-Thorsson G, Godaly G Mycobacteria bypass mucosal NF-kB signalling to induce an epithelial anti-inflammatory IL-22 and IL-10 response. PLoS ONE, 2014-01-28;9(1):e86466. 2014-01-28 [PMID: 24489729] (Western Blot, Human)
• Kitase Y, Barragan L, Qing H, Kondoh S, Jiang JX, Johnson ML, Bonewald LF Mechanical induction of PGE2 in osteocytes blocks glucocorticoid-induced apoptosis through both the beta-catenin and PKA pathways. J. Bone Miner. Res., 2010-06-24;25(12):2381-92. 2010-06-24 [PMID: 20578217] (Western Blot, Mouse)
• Tang Z, Arjunan P, Lee C, Li Y, Kumar A, Hou X, Wang B, Wardega P, Zhang F, Dong L, Zhang Y, Zhang SZ, Ding H, Fariss RN, Becker KG, Lennartsson J, Nagai N, Cao Y, Li X Survival effect of PDGF-CC rescues neurons from apoptosis in both brain and retina by regulating GSK3beta phosphorylation. J. Exp. Med., 2010-03-15;207(4):867-80. 2010-03-15 [PMID: 20231377] (Western Blot, Rat)

Bioinformatics

• Gene Symbol: GSK3A