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Human/Mouse/Rat Phospho-GSK-3 alpha / beta (S21/S9) Antibody

Catalog #: AF1590
6 Citations 1 Review Datasheet / COA / SDS
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AF1590
AF1590-SP
Detection of Human Phospho-GSK‑3 alpha (S21) and GSK-3 beta (S9) by Western Blot.
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Citations (6)
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Reviews (1)
Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human/Mouse/Rat Phospho-GSK-3 alpha / beta (S21/S9) Antibody Summary

Species Reactivity
Human, Mouse, Rat
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
Polyclonal Rabbit IgG
Purification
Antigen and protein A Affinity-purified
Immunogen
Phosphopeptide containing GSK-3 beta S9 site
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
0.2 µg/mL
See below
CyTOF-ready
Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
 
Immunocytochemistry
5-15 µg/mL
Immersion fixed HeLa human cervical epithelial carcinoma cell treated with PMA
Intracellular Staining by Flow Cytometry
2.5 µg/106 cells
See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot Detection of Human Phospho-GSK-3a (S21) and GSK-3 beta (S9) antibody by Western Blot. View Larger

Detection of Human Phospho-GSK‑3 alpha (S21) and GSK-3 beta (S9) by Western Blot. 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 (Catalog # 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 (Catalog # 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.

Immunocytochemistry View Larger

Phospho-GSK‑3 alpha / beta (S21/S9) in HeLa Human Cell Line. 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.

Intracellular Staining by Flow Cytometry Detection of Phospho-GSK-3a/ beta (S21/S9) antibody in Jurkat Human Cell Line antibody by Flow Cytometry. View Larger

Detection of Phospho-GSK‑3 alpha / beta (S21/S9) in Jurkat Human Cell Line by Flow Cytometry. Jurkat human acute T cell leukemia cells, untreated (open histogram, yellow line), or treated with PI3-kinase inhibitors (dark orange filled histogram), were stained with Human/Mouse/Rat Phospho-GSK-3a/ beta (S21/S9) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1590) or control antibody (Catalog # AB-105-C, blue open histogram), followed by Phycoerythrin-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # F0110). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
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Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & 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.

Background: GSK-3 alpha/beta

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.

Long Name
Glycogen Synthase Kinase 3
Alternate Names
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

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Citations for Human/Mouse/Rat Phospho-GSK-3 alpha / beta (S21/S9) Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

6 Citations: Showing 1 - 6
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  1. Multiscale networks in multiple sclerosis
    Authors: 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;
    PLoS computational biology
    Species: Human hepegivirus
    Sample Types: Whole Cells
    Applications: Flow Cytometry
  2. Chronic colitis upregulates microRNAs suppressing brain-derived neurotrophic factor in the adult heart
    Authors: Y Tang, KT Kline, XS Zhong, Y Xiao, H Lian, J Peng, X Liu, DW Powell, G Tang, Q Li
    PLoS ONE, 2021-09-20;16(9):e0257280.
    Species: Rat
    Sample Types: Tissue Homogenates
    Applications: Western Blot
  3. HYS-32-Induced Microtubule Catastrophes in Rat Astrocytes Involves the PI3K-GSK3beta Signaling Pathway.
    Authors: Chiu C, Liao C, Shen C, Tang T, Jow G, Wang H, Wu J
    PLoS ONE, 2015-05-04;10(5):e0126217.
    Species: Rat
    Sample Types: Cell Lysates
    Applications: Western Blot
  4. Mycobacteria bypass mucosal NF-kB signalling to induce an epithelial anti-inflammatory IL-22 and IL-10 response.
    Authors: Lutay N, Hakansson G, Alaridah N, Hallgren O, Westergren-Thorsson G, Godaly G
    PLoS ONE, 2014-01-28;9(1):e86466.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  5. Mechanical induction of PGE2 in osteocytes blocks glucocorticoid-induced apoptosis through both the beta-catenin and PKA pathways.
    Authors: Kitase Y, Barragan L, Qing H, Kondoh S, Jiang JX, Johnson ML, Bonewald LF
    J. Bone Miner. Res., 2010-06-24;25(12):2381-92.
    Species: Mouse
    Sample Types: Cell Lysates
    Applications: Western Blot
  6. Survival effect of PDGF-CC rescues neurons from apoptosis in both brain and retina by regulating GSK3beta phosphorylation.
    Authors: 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
    J. Exp. Med., 2010-03-15;207(4):867-80.
    Species: Rat
    Sample Types: Cell Lysates
    Applications: Western Blot

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Reviews for Human/Mouse/Rat Phospho-GSK-3 alpha / beta (S21/S9) Antibody

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Human/Mouse/Rat Phospho-GSK-3 alpha / beta (S21/S9) Antibody
By Kurt Patterson on 09/14/2018
Application: WB Sample Tested: Colon cancer cell line Species: Human
Western Blot Human/Mouse/Rat Phospho-GSK-3 alpha / beta (S21/S9) Antibody AF1590