GSK3A (Ab-21) Antibody
Product Specs
Target Background
GSK3A plays a significant role in insulin regulation of glycogen synthesis by phosphorylating and inhibiting GYS1 activity, thereby suppressing glycogen synthesis. It primarily regulates glycogen metabolism in the liver, but not in muscle. Additionally, GSK3A may contribute to the development of insulin resistance by modulating the activation of transcription factors.
Within the Wnt signaling pathway, GSK3A controls the levels and transcriptional activity of nuclear CTNNB1/beta-catenin. Furthermore, GSK3A facilitates the processing of amyloid precursor protein (APP) and the generation of APP-derived amyloid plaques, which are characteristic of Alzheimer's disease.
GSK3A may also be involved in the regulation of replication in pancreatic beta-cells. It is essential for establishing neuronal polarity and axon outgrowth. Through phosphorylation of the anti-apoptotic protein MCL1, GSK3A potentially regulates cell apoptosis in response to growth factor deprivation.
GSK3A acts as a regulator of autophagy by mediating the phosphorylation of KAT5/TIP60 under starvation conditions. This phosphorylation activates KAT5/TIP60 acetyltransferase activity, leading to the acetylation of crucial autophagy regulators, such as ULK1 and RUBCNL/Pacer.
Finally, GSK3A negatively regulates the extrinsic apoptotic signaling pathway mediated by death domain receptors. It promotes the formation of an anti-apoptotic complex, composed 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.
- GSK-3a and GSK-3b play pivotal roles in hepatocellular carcinoma. (Review) PMID: 28619606
- The combined action of CHIR99021 and forskolin markedly delayed neurological deficits and significantly reduced the tumor volume. Our findings suggest that reprogramming technology may be a potential treatment strategy replacing the traditional therapeutic paradigm of malignant glioma. Moreover, a combination molecule comprising a GSK3 inhibitor and a cAMP inducer could be the next generation of anticancer drugs. PMID: 29161257
- Using integrated analysis of genome-wide short hairpin RNA (shRNA) screening data in combination with genome-wide gene expression data, the study identified GSK3 as one of the key factors in p53-mediated apoptosis in human lung cancer cells. PMID: 28662516
- This study demonstrated that GSK-3alpha is regulated by CREB in lung cancer and is required for cell viability. These findings implicate the CREB-GSK-3alpha axis as a novel therapeutic target for lung cancer treatment. PMID: 27049759
- The study reveals that GSK3 acts through APC motifs R2 and B to regulate APC:Axin interactions, promoting high-throughput of betacatenin to destruction. PMID: 26393419
- This study reveals that GSK-3alpha- and GSK-3beta-regulated pathways can be responsible for stepwise transition to myelodysplastic syndromes and subsequent acute myeloid leukemia. PMID: 26766591
- CHP3 functions as a novel negative regulator of cardiomyocyte hypertrophy via inhibition of GSK3alpha/beta phosphorylation. PMID: 25935310
- Elevated GSK3 protein kinase activity is associated with non-small cell lung carcinoma. PMID: 25486534
- GSK3A is redundant with GSK3B in regulating drug-resistance and chemotherapy-induced necroptosis. PMID: 24984063
- ER stress-PERK-GSK3alpha/beta signaling promotes proatherogenic macrophage lipid accumulation. PMID: 25183803
- Glycogen synthase kinase 3 alpha and beta activity is increased in foetal membranes after term and preterm labour. PMID: 25550525
- TRAIL-induced apoptosis in pancreatic ductal adenocarcinoma cell lines is enhanced by pharmacological inhibition of glycogen synthase kinase-3 (GSK-3) or by shRNA-mediated depletion of either GSK-3alpha or GSK-3beta. PMID: 24675460
- Data indicate that autophagy inhibition promotes glycogen synthase kinase-(GSK3) inhibition-induced apoptosis. PMID: 25561726
- PtpA has a role in promoting anti-apoptotic activity of protein-dependent dephosphorylation of host GSK3alpha. PMID: 25187516
- Protein Kinase B-dependent phosphorylation of glycogen synthase kinase GSK-3 contributes to the pro-fibrotic signaling leading to renal tissue fibrosis in obstructive nephropathy. PMID: 24685987
- This review highlights the emerging role of GSK3alpha in Alzheimer's disease which has been largely overlooked in favor of GSK3beta. PMID: 24254703
- In lipopolysaccharide-stimulated human neutrophils, GSK-3-alpha inhibitors mimicked the effect of LiCl, by potentiating TNF-alpha release after 4 h. GSK-3 inhibition prolongs activation of the p38/MNK1/eIF4E pathway of mRNA translation. PMID: 24803542
- JNK and GSK3, in particular, are potential signaling molecules regulating BMP-induced osteoblastic human mesenchymal stem cells differentiation. PMID: 24060635
- The expression and regulation of GSK-3 in human neutrophils was investigated. PMID: 24035907
- This study demonstrated abundant levels of the critical negative cell-cycle regulators, p27(Kip1), its phosphorylated form, p-p27(S10), p18Ink4c, and GSK-3, in beta-cells of both adult human and mouse pancreatic islets, which contribute to the maintenance of beta-cell quiescence. PMID: 23896637
- The involvement of GSK-3 in several key pathophysiological pathways leading to AD and neurodegenerative diseases has placed this enzyme in a central position in this disorder. PMID: 23621307
- GSK3alpha, GSK3beta, and MAPK13 were found to be the most active tau kinases, phosphorylating tau at all four epitopes. PMID: 23798682
- Studies indicate the therapeutic potential of GSK-3 inhibitors for the treatment of neurodegenerative and behavior disorders. PMID: 23369789
- Lower phosphorylation levels of GSK3A are associated with a poor treatment response in rectal cancer. PMID: 22658458
- Findings suggest that inhibition of Bcl-2, Bcl-xL, and PI3K, and release of Bim from Bcl-2/Bcl-xL and GSK3alpha/beta culminating in Bax/Bak activation and apoptosis. PMID: 23243017
- GSK-3 may play a crucial role in the inflammatory process in chronic rhinosinusitis with nasal polyps (CRSwNP). PMID: 23352529
- GSK-3alpha is more prevalent in low Gleason score tumors, where it might promote androgen-dependent proliferation by activating the androgen receptor. PMID: 22539113
- Glycogen synthase kinase-3 (GSK3), when deinhibited by default in cells deprived of growth factors, activates acetyltransferase TIP60 through phosphorylating TIP60-Ser86, which acetylates and stimulates the protein kinase ULK1, which is required for autophagy. PMID: 22539723
- This study investigated whether GSK-3 activity is regulated by thymosin beta-4 (Tbeta4) and associated with Tbeta4-mediated migration in gastric cancer cells. Expression of GSK-3alpha, beta-catenin, and E-cadherin could be negatively regulated by Tbeta4-induced ERK phosphorylation. These findings suggest that Tbeta4 could be a novel regulator of the Wnt signaling pathway. PMID: 22328534
- The GSK-3 inhibitor BIO regulates proliferation of immortalized pancreatic mesenchymal stem cells. PMID: 22384031
- GSK-3alpha has a role and is a target in human acute myeloid leukemia. PMID: 22326953
- Glycogen synthase kinase (GSK)-3 promotes p70 ribosomal protein S6 kinase (p70S6K) activity and cell proliferation. PMID: 22065737
- Data show that p53 directly up-regulates microRNA-149* (miR-149*) that in turn targets glycogen synthase kinase-3alpha, resulting in increased expression of Mcl-1 and resistance to apoptosis in melanoma cells. PMID: 21896753
- GSK3 acts as a molecular brake on the signaling pathway, leading to TF expression in monocytes interacting with activated platelets. PMID: 21320285
- The authors demonstrate that inhibition of GSK-3 dampens Bacillus anthracis edema toxin-induced maturation and migration processes of monocyte-derived dendritic cells. PMID: 21576335
- Overexpression of glycogen synthase kinase-3 alpha in ovarian carcinoma cells is associated with acquired paclitaxel resistance. PMID: 21436692
- Investigations with human tissues in this review strongly support the postulate that GSK3 has a pathological role in mood disorders and is likely a therapeutic target in mood disorder treatment. PMID: 20668436
- Data show that the effects of PI3K-Akt signalling on IL-10 responses were mediated at least in part by GSK3. PMID: 21255011
- In multiple myeloma cells, GSK-3alpha and beta play distinct roles in cell survival and modulate the sensitivity to proteasome inhibitors. PMID: 20920357
- Phosphorylation of proteins in the brain associated with Alzheimer's disease is altered in cortical tissue lacking transgenic glycogen synthase kinase-3alpha and -3ss. PMID: 20831597
- GSK3A is widely expressed throughout the hair follicles. PMID: 19705135
- Glycogen synthase kinase (GSK3) plays a central role in promoting glucocorticoid-induced apoptosis. PMID: 20371704
- GSK-3 maintains the MLL leukemia stem cell transcriptional program by promoting the conditional association of CREB and its coactivators TORC and CBP with homedomain protein MEIS1, which in turn facilitates HOX-mediated transcription and transformation. PMID: 20541704
- This study suggests that downstream signaling components of the PI3K/Akt pathway, GSK3 & cyclin D2, as well as the significant interaction between PTEN-PDK and between pAkt-pGSK3beta, are involved in the survival and proliferation of leiomyomas. PMID: 19464003
- The N termini of both GSK-3 isoforms were dispensable, whereas progressive C-terminal deletions resulted in protein misfolding exhibited by deficient activity, impaired ability to interact with Axin, and a loss of Tyr(279/216) phosphorylation. PMID: 20080974
- Studies show that GSK-3 and Omi/HtrA2 synergistically cause annexin A2 cleavage and then cell cycle inhibition or apoptosis. PMID: 19656851
- Immunoreactivity of IGF-I and GSK3a/b signaling pathways was studied in ALS spinal cords and hippocampus with special reference to Kii and Guam ALS patients. In Japanese ALS patients, IGF-I expression was positively correlated with the clinical duration. PMID: 19323791
- GSK3A improves insulin action and glucose metabolism in skeletal muscle. PMID: 12086949
- GSK3alpha is a potential regulator of platelet function. PMID: 14550568
- Data show that myotubes defective in glycogen synthase (GS) activity express insulin-responsive glycogen synthase kinase-3alpha, suggesting that failure of insulin to decrease GS phosphorylation involves abnormal activity of another kinase or phosphatase. PMID: 15194499
Q&A
What is GSK3A (Ab-21) Antibody and what epitope does it recognize?
GSK3A (Ab-21) Antibody is a rabbit polyclonal antibody that specifically recognizes the glycogen synthase kinase-3 alpha isoform. It was generated by immunizing rabbits with a synthetic peptide corresponding to amino acids 19-23 (T-S-S-F-A) of human GSK3α conjugated to KLH (keyhole limpet hemocyanin) . This antibody detects endogenous levels of total GSK3α protein and demonstrates high specificity, not cross-reacting with the GSK3β isoform .
What are the validated applications for GSK3A (Ab-21) Antibody?
The GSK3A (Ab-21) Antibody has been validated for multiple experimental applications:
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Western Blotting (WB): Recommended dilution of 1:500-1:1000
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Immunohistochemistry (IHC): Recommended dilution of 1:50-1:100
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Immunofluorescence (IF): Recommended dilution of 1:100-1:200
Western blot analysis has demonstrated successful detection of GSK3α in cell extracts from 293 cells, with the antibody detecting a band at the predicted molecular weight of 51 kDa .
How specific is GSK3A (Ab-21) Antibody, and how does it differ from other GSK3 antibodies?
GSK3A (Ab-21) Antibody exhibits high specificity for GSK3α and does not detect GSK3β . This is particularly important given the structural similarities between the two isoforms. In contrast, some antibodies like clone 15C2 recognize both GSK3α and GSK3β, specifically in their non-phosphorylated forms at S21 and S9 respectively . The GSK3A (Ab-21) Antibody detects total GSK3α regardless of its phosphorylation status at S21, making it distinct from phospho-specific antibodies or non-phospho-specific antibodies like 12B2 and 15C2 .
What is the optimal storage condition for preserving GSK3A (Ab-21) Antibody activity?
For long-term preservation, it is recommended to store the GSK3A (Ab-21) Antibody at -20°C. The antibody is supplied at a concentration of 1.0 mg/mL in phosphate buffered saline (without Mg2+ and Ca2+), pH 7.4, with 150 mM NaCl, 0.02% sodium azide, and 50% glycerol . For short-term use, the antibody can be stored at 4°C, though prolonged storage at this temperature may compromise antibody performance .
What controls should be included when using GSK3A (Ab-21) Antibody in experiments?
When designing experiments with GSK3A (Ab-21) Antibody, researchers should consider the following controls:
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Negative control: Preincubation of the antibody with its specific blocking peptide, which should eliminate the signal in applications like IHC, as demonstrated in human breast carcinoma tissue .
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Specificity control: Including GSK3β samples to verify the lack of cross-reactivity reported for this antibody .
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Loading control: Using a validated housekeeping protein antibody in parallel for normalization in Western blot applications.
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Positive control: Cell lines known to express GSK3α, such as 293 cells, HeLa, or MCF-7 human cells, which have been successfully used in validation studies .
How should samples be prepared for optimal detection with GSK3A (Ab-21) Antibody?
For Western blotting:
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Prepare cell lysates using standard protocols with protease inhibitors to prevent protein degradation
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Use reducing conditions for SDS-PAGE as demonstrated in validation studies
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Transfer proteins to PVDF membrane for optimal antibody binding
For Immunohistochemistry:
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Paraffin-embedded tissues have been successfully used with this antibody
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Follow standard antigen retrieval protocols appropriate for formalin-fixed tissues
For Immunofluorescence:
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Methanol fixation has been validated for cell lines such as MCF7
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For intracellular staining in flow cytometry applications, similar to other GSK3 antibodies, paraformaldehyde fixation and saponin permeabilization may be used
How can GSK3A (Ab-21) Antibody be used to study GSK3α regulatory mechanisms?
GSK3A (Ab-21) Antibody can be valuable for studying GSK3α regulation by detecting total GSK3α levels independent of its phosphorylation state. When used in conjunction with phospho-specific antibodies that detect GSK3α phosphorylated at S21, researchers can assess the proportion of active versus inactive GSK3α in various experimental conditions .
A notable research strategy involves using this antibody alongside non-phosphorylated serine-specific antibodies like 12B2 (specific for non-phospho-S9 GSK3β) and 15C2 (specific for non-phospho-S9/21 GSK3β/α). This approach allows for comprehensive analysis of GSK3 regulation by:
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Measuring total GSK3α levels with GSK3A (Ab-21) Antibody
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Assessing active GSK3α (non-phosphorylated at S21) with non-phospho-specific antibodies
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Quantifying inactive GSK3α (phosphorylated at S21) with phospho-specific antibodies
How does phosphorylation affect GSK3α function and antibody recognition?
Phosphorylation of GSK3α at serine 21 serves as a primary regulatory mechanism that inhibits its kinase activity. When S21 is phosphorylated, the N-terminus acts as a competitive inhibitor for primed substrates, thereby reducing GSK3α activity . The GSK3A (Ab-21) Antibody recognizes an epitope (aa.19-23) that is in close proximity to this critical regulatory site.
What experimental approaches can be used to study GSK3α and GSK3β isoform-specific functions?
The two GSK3 isoforms (GSK3α and GSK3β) have distinctive functions and are not redundant, as evidenced by their different knockout phenotypes and regulatory roles . The GSK3A (Ab-21) Antibody's specificity for GSK3α makes it an excellent tool for studying isoform-specific functions through several approaches:
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Comparative expression analysis: Using GSK3A (Ab-21) Antibody alongside GSK3β-specific antibodies to compare expression patterns across tissues or cellular compartments .
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Knockdown/knockout validation: Confirming the specificity of gene silencing or knockout strategies for GSK3α by demonstrating the absence of signal with GSK3A (Ab-21) Antibody while GSK3β signals remain intact .
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Isoform-specific interactions: Employing GSK3A (Ab-21) Antibody in co-immunoprecipitation studies to identify proteins that interact specifically with GSK3α but not GSK3β.
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Regulatory pathway analysis: Investigating how different signaling pathways might differentially regulate GSK3α versus GSK3β by monitoring changes in GSK3α levels and phosphorylation states in response to various stimuli .
What technical considerations should be addressed when analyzing GSK3α in complex biological systems?
When using GSK3A (Ab-21) Antibody to study GSK3α in complex biological systems, researchers should consider:
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Isoform discrimination: While GSK3A (Ab-21) Antibody is reported to be specific for GSK3α, validation in the specific biological system under study is recommended, particularly in tissues with high expression of both isoforms .
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Phosphorylation dynamics: When studying GSK3α activity, remember that GSK3A (Ab-21) Antibody detects total GSK3α regardless of phosphorylation state. Consider parallel analysis with phospho-specific antibodies to determine active versus inactive proportions .
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Subcellular localization: GSK3α has been observed in both cytoplasmic and nuclear compartments. When using GSK3A (Ab-21) Antibody for localization studies, appropriate subcellular fractionation or high-resolution imaging techniques should be employed .
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Species considerations: While GSK3A (Ab-21) Antibody has reactivity to human, mouse, and rat GSK3α, sequence variations across more distant species may affect epitope recognition, necessitating validation in non-validated species .
What are common issues when using GSK3A (Ab-21) Antibody and how can they be resolved?
When working with GSK3A (Ab-21) Antibody, researchers might encounter several common issues:
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Low signal in Western blotting:
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Optimize antibody concentration (try 1:500 dilution if 1:1000 yields weak signals)
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Increase protein loading (GSK3α is expressed at moderate levels in many cell types)
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Extend exposure time or use more sensitive detection methods
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Ensure transfer efficiency by using protein standards or reversible stains
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Background in immunohistochemistry:
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Weak immunofluorescence signal:
How does GSK3A (Ab-21) Antibody perform compared to other commercially available GSK3α antibodies?
The GSK3A (Ab-21) Antibody has distinct advantages compared to other GSK3α antibodies:
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Isoform specificity: Unlike some total GSK3 antibodies that recognize both α and β isoforms, GSK3A (Ab-21) Antibody is specific for GSK3α .
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Multiple application validation: It has been validated for Western blotting, immunohistochemistry, and immunofluorescence, making it versatile for various experimental approaches .
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Targeted epitope: The antibody recognizes an N-terminal epitope (aa.19-23) that is distinct from the phosphorylation site (S21), allowing for detection of total GSK3α regardless of activation state .
In contrast, antibodies like clone 12B2 (specific for non-phospho-S9 GSK3β) and clone 15C2 (specific for non-phospho-S9/21 GSK3β/α) are designed specifically to distinguish phosphorylation states, which serves a different experimental purpose than GSK3A (Ab-21) Antibody .
How is GSK3α involved in disease pathways, and how can GSK3A (Ab-21) Antibody contribute to such research?
GSK3α has been implicated in various disease processes, and GSK3A (Ab-21) Antibody can be instrumental in investigating these associations:
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Cancer: GSK3 has been shown to play both tumor suppressor and promoter roles depending on context. In some cases, GSK3 controls the expression of PD-1 and can serve as a tumor promoter . GSK3A (Ab-21) Antibody has been validated in cancer cell lines including HeLa cervical carcinoma, HT-29 colon adenocarcinoma, and MCF-7 breast cancer cells , making it useful for studying GSK3α's role in cancer biology.
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Neurodegenerative diseases: GSK3 isoforms are involved in tau phosphorylation and amyloid processing. GSK3A (Ab-21) Antibody can help distinguish isoform-specific contributions to these pathological processes .
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Metabolic disorders: Given GSK3's role in glycogen metabolism, GSK3A (Ab-21) Antibody can be used to study how GSK3α levels or localization might be altered in metabolic syndrome or diabetes.
How can GSK3A (Ab-21) Antibody be used in studies involving GSK3 inhibitors and therapeutic approaches?
The GSK3A (Ab-21) Antibody can be valuable in studies involving GSK3 inhibitors and therapeutic approaches:
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Inhibitor specificity: When evaluating GSK3 inhibitors such as SB216763, GSK inhibitor XIII, or AR-A014418, researchers can use GSK3A (Ab-21) Antibody to determine whether these compounds affect total GSK3α levels in addition to their effects on kinase activity .
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Target validation: In approaches targeting GSK3α specifically (versus GSK3β or both isoforms), GSK3A (Ab-21) Antibody can confirm target engagement or knockdown efficiency.
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Biomarker development: GSK3A (Ab-21) Antibody might be useful in evaluating GSK3α as a potential biomarker in diseases where GSK3 dysregulation is implicated, such as certain cancers or neurodegenerative disorders.
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Mechanism studies: When investigating how compounds like the androgen receptor (AR) inhibitors affect GSK3 signaling pathways, GSK3A (Ab-21) Antibody can help elucidate isoform-specific mechanisms of action .
