Recombinant Anti-GSK3 beta (phospho S9) antibody [EPR2286Y] (ab75814)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR2286Y] to GSK3 beta (phospho S9)
- Suitable for: Dot blot, WB, IHC-P, ICC/IF
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-GSK3 beta (phospho S9) antibody [EPR2286Y]
See all GSK3 beta primary antibodies -
Description
Rabbit monoclonal [EPR2286Y] to GSK3 beta (phospho S9) -
Host species
Rabbit -
Tested applications
Suitable for: Dot blot, WB, IHC-P, ICC/IFmore details
Unsuitable for: Flow Cyt or IP -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- 293T cell lysates, untreated or treated with Calyculin A and human ovarian carcinoma tissue.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR2286Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Positive Controls
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab75814 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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Dot blot |
Use at an assay dependent concentration.
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WB | (7) |
1/10000 - 1/20000. Detects a band of approximately 47 kDa (predicted molecular weight: 47 kDa).
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IHC-P | (1) |
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
|
ICC/IF | (2) |
1/100 - 1/250.
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Notes |
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Dot blot
Use at an assay dependent concentration. |
WB
1/10000 - 1/20000. Detects a band of approximately 47 kDa (predicted molecular weight: 47 kDa). |
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
ICC/IF
1/100 - 1/250. |
Target
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Function
Participates in the Wnt signaling pathway. Implicated in the hormonal control of several regulatory proteins including glycogen synthase, MYB and the transcription factor JUN. Phosphorylates JUN at sites proximal to its DNA-binding domain, thereby reducing its affinity for DNA. Phosphorylates MUC1 in breast cancer cells, and decreases the interaction of MUC1 with CTNNB1/beta-catenin. Phosphorylates CTNNB1/beta-catenin. Phosphorylates SNAI1. Plays an important role in ERBB2-dependent stabilization of microtubules at the cell cortex. Prevents the phosphorylation of APC and CLASP2, allowing its association with the cell membrane. In turn, membrane-bound APC allows the localization of MACF1 to the cell membrane, which is required for microtubule capture and stabilization. Phosphorylates MACF1 and this phosphorylation inhibits the binding of MACF1 to microtubules which is critical for its role in bulge stem cell migration and skin wound repair. -
Tissue specificity
Expressed in testis, thymus, prostate and ovary and weakly expressed in lung, brain and kidney. -
Sequence similarities
Belongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. GSK-3 subfamily.
Contains 1 protein kinase domain. -
Post-translational
modificationsPhosphorylated by AKT1 and ILK1. Activated by phosphorylation at Tyr-216. -
Cellular localization
Cytoplasm. Nucleus. Cell membrane. The phosphorylated form shows localization to cytoplasm and cell membrane. The MEMO1-RHOA-DIAPH1 signaling pathway controls localization of the phosophorylated form to the cell membrane. - Information by UniProt
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Database links
- Entrez Gene: 2932 Human
- Omim: 605004 Human
- SwissProt: P49841 Human
- Unigene: 445733 Human
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Alternative names
- Glycogen Synthase Kinase 3 Beta antibody
- Glycogen synthase kinase-3 beta antibody
- GSK 3 beta antibody
see all
Images
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All lanes : Anti-GSK3 beta (phospho S9) antibody [EPR2286Y] (ab75814) at 1/20000 dilution
Lane 1 : Untreated HeLa cell lysate
Lane 2 : HeLa cells were starved for 3 hours then treated with Calyculin A (100nM) for 30 minutes.
Lane 3 : HeLa cells were starved for 3 hours then treated with Calyculin A (100nM) for 30 minutes. The membrane was treated with alkaline phosphatase.
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat anti-rabbit IgG H&L (HRP))
Predicted band size: 47 kDa
Observed band size: 47 kDa
Exposure time: 5 secondsBlocking buffer: 5% NFDM/TBST for the first 3 lanes, 2% BSA/TBST for the second 3 lanes.
Dilution buffer: 5% NFDM/TBST for the first 3 lanes, 2% BSA/TBST for the second 3 lanes.
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Cell line: HeLa (human cervix adenocarcinoma)
Target: Ab75814 anti-GSK3 beta (phospho S9)
Secondary: ab150077 AlexaFluor®488 Goat anti-Rabbit
Fixative: 4% PFA
Permeabilization: 0.1% Triton-X
Nuclear counter stain: DAPI
Confocal image showed increased cytoplasmic staining after Calyculin A (CA) (ab141784 100ng/ml, 10min) treatment on Hela cells. The LP treatment decreased the increased cytoplasmic staining caused by CA.
Ab32391 was used as a control for ab75814. The results showed cytoplasmic staining on untreated, CA and CA+LP treated Hela cells.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSK3 beta (phospho S9) antibody [EPR2286Y] (ab75814)
Immunohistochemical analysis of paraffin-embedded human breast tissue labelled with untreated ab75814 (phospho) (top-left) at a dilution of 1/1000, alkaline phosphatase treated ab75814 (phospho) (top-right) at a dilution of 1/1000, untreated ab32391 (bottom-left) at a dilution of 1/1000 and alkaline phophatase treated ab32391 (bottom-right) at a dilution of 1/1000. Ab97051 was used as secondary antibody at a dilution of 1/500 and counterstained with hematoxylin.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Dot blot analysis of GSK3 beta (pS9) peptide (Lane 1) and GSK3 beta non-phospho peptide (Lane 2) labelling GSK3 (pS9) with purified ab75814 at a dilution of 1/1000. ab97051 (Peroxidase conjugated goat anti-rabbit IgG (H+L)) was used as the secondary antibody at a dilution of 1/100000.
Blocking and dilution buffer: 5% NFDM/TBST.
Exposure time: 3 minutes.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSK3 beta (phospho S9) antibody [EPR2286Y] (ab75814)
Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labelled with untreated ab75814 (top-left) at a dilution of 1/1000, alkaline phosphatase treated ab75814 (top-right) at a dilution of 1/1000, untreated ab32391 (bottom-left) at a dilution of 1/1000 and alkaline phophatase treated ab32391 (bottom-right) at a dilution of 1/1000. Ab97051 was used as secondary antibody at a dilution of 1/500 and counterstained with hematoxylin.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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All lanes : Anti-GSK3 beta (phospho S9) antibody [EPR2286Y] (ab75814) at 1/20000 dilution
Lane 1 : 293T cell lysates, untreated
Lane 2 : 293T cell lysates, treated with Calyculin A
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : goat anti-rabbit HRP at 1/2000 dilution
Predicted band size: 47 kDa
Observed band size: 47 kDa
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (100)
ab75814 has been referenced in 100 publications.
- Pei X et al. The neuroprotective effects of alpha-lipoic acid on an experimental model of Alzheimer's disease in PC12 cells. J Appl Toxicol 42:285-294 (2022). PubMed: 34133789
- Boyle AJ et al. PET imaging of glycogen synthase kinase-3 in pancreatic cancer xenograft mouse models. Am J Nucl Med Mol Imaging 12:1-14 (2022). PubMed: 35295885
- Lei T et al. Panax notoginseng saponins improves healing of high glucose-induced wound through the GSK-3β/β-catenin pathway. Environ Toxicol 37:1867-1877 (2022). PubMed: 35385194
- Zhao Q et al. Visfatin inhibits colon cancer cell apoptosis and decreases chemosensitivity to 5‑FU by promoting the SDF‑1/CXCR4/Akt axis. Int J Oncol 60:N/A (2022). PubMed: 35506454
- Sun X et al. PRDX2 Knockdown Inhibits Extracellular Matrix Synthesis of Chondrocytes by Inhibiting Wnt5a/YAP1/CTGF and Activating IL-6/JAK2/STAT3 Pathways in Deer Antler. Int J Mol Sci 23:N/A (2022). PubMed: 35563622