Name: Anti-GSK3 beta antibody [Y174]
SKU: ab32391
Rating: 5 (2 reviews)

Rabbit Recombinant Monoclonal GSK3 beta antibody. Suitable for IHC-P, ELISA, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human, Synthetic peptide - Human samples. Cited in 206 publications.

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Images

Western blot - Anti-GSK3 beta antibody [Y174] (AB32391), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	IHC-P	ELISA	WB	ICC/IF	Flow Cyt (Intra)
Human	Tested	Expected	Tested	Tested	Tested
Mouse	Predicted	Predicted	Predicted	Predicted	Predicted
Synthetic peptide - Human	Not recommended	Tested	Not recommended	Not recommended	Not recommended

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Predicted
Predicted

Species	Dilution info	Notes
Species Mouse	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Synthetic peptide - Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Synthetic peptide - Human	Dilution info -	Notes Unit Type: 0 - 1000 ng/ml

Expected
Expected

Species	Dilution info	Notes
Species Human	Dilution info Use at an assay dependent concentration.	Notes -

Predicted
Predicted

Species	Dilution info	Notes
Species Mouse	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/5000 - 1/10000	Notes -

Predicted
Predicted

Species	Dilution info	Notes
Species Mouse	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Synthetic peptide - Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/100 - 1/500	Notes -

Predicted
Predicted

Species	Dilution info	Notes
Species Mouse	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Synthetic peptide - Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/100	Notes ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Predicted
Predicted

Species	Dilution info	Notes
Species Mouse	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Synthetic peptide - Human	Dilution info -	Notes -

Associated Products

Select an associated product type

9 products for Alternative Product

Target data

See full target information GSK3B

Function

Constitutively active protein kinase that acts as a negative regulator in the hormonal control of glucose homeostasis, Wnt signaling and regulation of transcription factors and microtubules, by phosphorylating and inactivating glycogen synthase (GYS1 or GYS2), EIF2B, CTNNB1/beta-catenin, APC, AXIN1, DPYSL2/CRMP2, JUN, NFATC1/NFATC, MAPT/TAU and MACF1 (PubMed:11430833, PubMed:12554650, PubMed:14690523, PubMed:16484495, PubMed:1846781, PubMed:20937854, PubMed:9072970). Requires primed phosphorylation of the majority of its substrates (PubMed:11430833, PubMed:16484495). In skeletal muscle, contributes to insulin regulation of glycogen synthesis by phosphorylating and inhibiting GYS1 activity and hence glycogen synthesis (PubMed:8397507). May also mediate the development of insulin resistance by regulating activation of transcription factors (PubMed:8397507). Regulates protein synthesis by controlling the activity of initiation factor 2B (EIF2BE/EIF2B5) in the same manner as glycogen synthase (PubMed:8397507). In Wnt signaling, GSK3B forms a multimeric complex with APC, AXIN1 and CTNNB1/beta-catenin and phosphorylates the N-terminus of CTNNB1 leading to its degradation mediated by ubiquitin/proteasomes (PubMed:12554650). Phosphorylates JUN at sites proximal to its DNA-binding domain, thereby reducing its affinity for DNA (PubMed:1846781). Phosphorylates NFATC1/NFATC on conserved serine residues promoting NFATC1/NFATC nuclear export, shutting off NFATC1/NFATC gene regulation, and thereby opposing the action of calcineurin (PubMed:9072970). Phosphorylates MAPT/TAU on 'Thr-548', decreasing significantly MAPT/TAU ability to bind and stabilize microtubules (PubMed:14690523). MAPT/TAU is the principal component of neurofibrillary tangles in Alzheimer disease (PubMed:14690523). Plays an important role in ERBB2-dependent stabilization of microtubules at the cell cortex (PubMed:20937854). Phosphorylates MACF1, inhibiting its binding to microtubules which is critical for its role in bulge stem cell migration and skin wound repair (By similarity). Probably regulates NF-kappa-B (NFKB1) at the transcriptional level and is required for the NF-kappa-B-mediated anti-apoptotic response to TNF-alpha (TNF/TNFA) (By similarity). Negatively regulates replication in pancreatic beta-cells, resulting in apoptosis, loss of beta-cells and diabetes (By similarity). Through phosphorylation of the anti-apoptotic protein MCL1, may control cell apoptosis in response to growth factors deprivation (By similarity). Phosphorylates MUC1 in breast cancer cells, decreasing the interaction of MUC1 with CTNNB1/beta-catenin (PubMed:9819408). Is necessary for the establishment of neuronal polarity and axon outgrowth (PubMed:20067585). Phosphorylates MARK2, leading to inhibition of its activity (By similarity). Phosphorylates SIK1 at 'Thr-182', leading to sustainment of its activity (PubMed:18348280). Phosphorylates ZC3HAV1 which enhances its antiviral activity (PubMed:22514281). Phosphorylates SNAI1, leading to its ubiquitination and proteasomal degradation (PubMed:15448698, PubMed:15647282, PubMed:25827072, PubMed:29059170). Phosphorylates SFPQ at 'Thr-687' upon T-cell activation (PubMed:20932480). Phosphorylates NR1D1 st 'Ser-55' and 'Ser-59' and stabilizes it by protecting it from proteasomal degradation. Regulates the circadian clock via phosphorylation of the major clock components including BMAL1, CLOCK and PER2 (PubMed:19946213, PubMed:28903391). Phosphorylates FBXL2 at 'Thr-404' and primes it for ubiquitination by the SCF(FBXO3) complex and proteasomal degradation (By similarity). Phosphorylates CLOCK AT 'Ser-427' and targets it for proteasomal degradation (PubMed:19946213). Phosphorylates BMAL1 at 'Ser-17' and 'Ser-21' and primes it for ubiquitination and proteasomal degradation (PubMed:28903391). Phosphorylates OGT at 'Ser-3' or 'Ser-4' which positively regulates its activity. Phosphorylates MYCN in neuroblastoma cells which may promote its degradation (PubMed:24391509). Regulates the circadian rhythmicity of hippocampal long-term potentiation and BMAL1 and PER2 expression (By similarity). Acts as a regulator of autophagy by mediating phosphorylation of KAT5/TIP60 under starvation conditions, activating KAT5/TIP60 acetyltransferase activity and promoting acetylation of key autophagy regulators, such as ULK1 and RUBCNL/Pacer (PubMed:30704899). Negatively regulates extrinsic apoptotic signaling pathway via death domain receptors. Promotes the formation of an anti-apoptotic complex, made of DDX3X, BRIC2 and GSK3B, at death receptors, including TNFRSF10B. The anti-apoptotic function is most effective with weak apoptotic signals and can be overcome by stronger stimulation (PubMed:18846110). Phosphorylates E2F1, promoting the interaction between E2F1 and USP11, stabilizing E2F1 and promoting its activity (PubMed:17050006, PubMed:28992046). Phosphorylates mTORC2 complex component RICTOR at 'Thr-1695' which facilitates FBXW7-mediated ubiquitination and subsequent degradation of RICTOR (PubMed:25897075). Phosphorylates FXR1, promoting FXR1 ubiquitination by the SCF(FBXO4) complex and FXR1 degradation by the proteasome (By similarity). Phosphorylates interleukin-22 receptor subunit IL22RA1, preventing its proteasomal degradation (By similarity).

Alternative names

Glycogen synthase kinase-3 beta, GSK-3 beta, Serine/threonine-protein kinase GSK3B, GSK3B

Recommended products

Rabbit Recombinant Monoclonal GSK3 beta antibody. Suitable for IHC-P, ELISA, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human, Synthetic peptide - Human samples. Cited in 206 publications.

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form

Liquid

Clonality

Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Clone number

Y174

Purification technique

Affinity purification Protein A

Specificity

This antibody is specific for human GSK3 beta. It may also detect the splice isoform 2 based on sequence homology.

The immunogen used for this antibody is GSK3 beta phospho S9. This antibody shows partially phospho specificity to phospho S9 under certain conditions, for example, under low peptide concentration in ELISA assay, it has dominant reactivity with phospho S9 peptide.

Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

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 batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

Glycogen synthase kinase 3 beta or GSK3 beta is a serine/threonine protein kinase that regulates various cellular processes. It is also referred to as GSK3B and has a molecular weight of approximately 47 kilodaltons (kDa). This enzyme is widely expressed in many tissues with notably high expression levels in the brain. Mechanically GSK3 beta phosphorylates target proteins affecting their activity stability and interactions with other proteins.

Biological function summary

As an important regulatory protein GSK3 beta plays a role in multiple cellular functions including metabolism cell cycle and cell signaling. It often interacts with components in signaling complexes and modulates the activity of these complexes involving phosphorylation. It is involved in the regulation of transcription factors such as c-Myc and β-catenin influencing cell fate and survival. This ability to modulate diverse functions highlights GSK3 beta's importance in cellular regulation.

Pathways

GSK3 beta participates in significant signaling pathways such as the Wnt/β-catenin and insulin signaling pathways. In the Wnt pathway GSK3 beta phosphorylates β-catenin marking it for degradation which regulates gene transcription. In insulin signaling it interacts with proteins like AKT to mediate glycogen synthesis. These interactions highlight its role in maintaining cellular homeostasis and energy balance.

Associated diseases and disorders

GSK3 beta has implications in Alzheimer's disease and type 2 diabetes. It is associated with tau protein hyperphosphorylation and neurofibrillary tangles in Alzheimer's disease. In diabetes its role in insulin signaling impacts glucose metabolism. Further GSK3 beta's dysfunction relates to disrupted signaling related to these conditions positioning it as a potential therapeutic target.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
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11 product images

Western blot - Anti-GSK3 beta antibody [Y174] (ab32391)
Lane 1 & 3: Wild type HAP1 whole cell lysate (20 μg)
Lane 2 & 4: GSK3B knockout HAP1 whole cell lysate (20 μg)
Lanes 1 - 4: Green - ab32391 observed at 46 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245, observed at 37 kDa.
ab32391 was shown to recognize GSK3B in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when GSK3B knockout samples were examined. Wild-type and GSK3B knockout samples were subjected to SDS-PAGE. ab32391 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/2000 dilution and 1/10,000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-GSK3 beta antibody [Y174] (ab32391)
Predicted band size: 46 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSK3 beta antibody [Y174] (ab32391)
Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labelled with untreated Anti-GSK3 beta (phospho S9) antibody [EPR2286Y] ab75814 (phospho) (top-left) at a dilution of 1/1000, alkaline phosphatase treated Anti-GSK3 beta (phospho S9) antibody [EPR2286Y] 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. Goat Anti-Rabbit IgG H&L (HRP) ab97051 was used as secondary antibody at a dilution of 1/500 and counterstained with hematoxylin.
Immunocytochemistry/ Immunofluorescence - Anti-GSK3 beta antibody [Y174] (ab32391)
Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling GSK3 beta with purified ab32391 at 1/500. Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% tritonX-100. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/1000 dilution was used as the secondary antibody. Cells were co-stained with Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, a mouse anti-tubulin antibody (1/1000) using Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) as the secondary antibody. Nuclei were counterstained with DAPI (blue).
For negative control 1, rabbit primary antibody was used followed by anti-mouse secondary antibody (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120).
For negative control 2, mouse primary antibody (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) and anti-rabbit secondary antibody (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) were used.
Western blot - Anti-GSK3 beta antibody [Y174] (ab32391)
All lanes: Western blot - Anti-GSK3 beta antibody [Y174] (ab32391) at 1/10000 dilution
All lanes: A431 cell lysate.
Predicted band size: 46 kDa
Observed band size: 46 kDa
Flow Cytometry (Intracellular) - Anti-GSK3 beta antibody [Y174] (ab32391)
Overlay histogram showing HeLa cells stained with ab32391 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32391, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.
Image courtesy of an anonymous customer review.
Western blot - Anti-GSK3 beta antibody [Y174] (ab32391)
All lanes: Western blot - Anti-GSK3 beta antibody [Y174] (ab32391) at 1/2500 dilution
Lane 1: MCF7 (human breast adenocarcinoma cell line) cell lysate at 20 µg
Lane 2: SW480 (human colorectal adenocarcinoma cell line) cell lysate at 20 µg
Secondary
All lanes: Donkey polyclonal IRDye 800CW at 1/15000 dilution
Predicted band size: 46 kDa
Observed band size: 37 kDa, 46 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSK3 beta antibody [Y174] (ab32391)
Immunohistochemical analysis of paraffin-embedded human breast tissue labelled with untreated Anti-GSK3 beta (phospho S9) antibody [EPR2286Y] ab75814 (phospho) (top-left) at a dilution of 1/1000, alkaline phosphatase treated Anti-GSK3 beta (phospho S9) antibody [EPR2286Y] 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. Goat Anti-Rabbit IgG H&L (HRP) ab97051 was used as secondary antibody at a dilution of 1/500 and counterstained with hematoxylin.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSK3 beta antibody [Y174] (ab32391)
IHC image of GSK3 staining in human breast adenocarcinoma formalin fixed paraffin embedded tissue section, performed on a Leica Bond system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6, epitope retrieval solution 1) for 20 minutes. The section was then incubated with ab32391, 1/200 diution, for 15 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
Immunocytochemistry/ Immunofluorescence - Anti-GSK3 beta antibody [Y174] (ab32391)
ICC/IF image of ab32391 stained DU145 cells. The cells were 4% formaldehyde fixed (10 minutes) and then incubated in 1 %BSA / 10 % normal goat serum / 0.3 M glycine in 0.1 % PBS-Tween for 1 hour to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab32391 at 1/200 dilution overnight at +4°C. The secondary antibody (green) was DyLight^® 488 goat anti- rabbit (Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed ab96899) IgG (H+L) used at a 1/250 dilution for 1 hour. Alexa Fluor^® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1 hour. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43 μM.
ELISA - Anti-GSK3 beta antibody [Y174] (ab32391)
ELISA of GSK3 beta (phospho S9) peptide and GSK3 beta non-phospho peptide at 10 ng/ml. Detected with ab32391 at 0~1000 ng/ml. Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1/2500 was used as a secondary antibody.
ELISA - Anti-GSK3 beta antibody [Y174] (ab32391)
ELISA of GSK3 beta (phospho S9) peptide and GSK3 beta non-phospho peptide at 1000 ng/ml. Detected with ab32391 at 0~1000 ng/ml. Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1/2500 was used as a secondary antibody.