Phospho-GSK3 beta (S9) Recombinant Rabbit Monoclonal Antibody [SY02-71] - BSA and Azide free
Catalog# HA750128
Phospho-GSK3 beta (S9) Recombinant Rabbit Monoclonal Antibody [SY02-71] - BSA and Azide free
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WB
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IF-Cell
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IF-Tissue
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IHC-P
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FC
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Human
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Mouse
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Rat
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ET1607-60
含抗保成分
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unconjugated
Overview
Product Name
Phospho-GSK3 beta (S9) Recombinant Rabbit Monoclonal Antibody [SY02-71] - BSA and Azide free
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic phospho-peptide corresponding to residues surrounding Ser9 of Human GSK3 beta aa 1-50 / 420.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IF-Cell, IF-Tissue, IHC-P, FC
Target Molecular Weight
Predicted band size: 47 kDa
Positive Control
HeLa treated with 100nM Calyculin A for 30 minutes cell lysate, NIH/3T3 cell lysate, NIH/3T3 treated with 100ng/mL PDGF for 5 minutes cell lysate, C6 cell lysate, C6 treated with 100ng/mL PDGF for 5 minutes cell lysate, C6 treated with 150nM insulin for 15 minutes cell lysate, HeLa cells treated with 100nM Calyculin A for 30 minutes, NIH/3T3 cells treated with 100ng/mL PDGF for 5 minutes, C6 cells treated with 100ng/mL PDGF for 5 minutes, human colon carcinoma tissue, human breast tissue, human breast carcinoma tissue, human kidney tissue, human pancreas tissue.
Conjugation
unconjugated
Clone Number
SY02-71
Product Features
Form
Liquid
Concentration
1mg/ml
Storage Instructions
Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage Buffer
1*PBS (pH7.4).
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:5,000-1:10,000
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IF-Cell
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1:100-1:500
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IF-Tissue
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1:50-1:200
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IHC-P
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1:50-1:200
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FC
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1:1,000
Target
Function
Glycogen synthase kinase-3α (GSK-3α) and GSK-3β are highly similar isoforms of serine/ threonine kinases that regulate metabolic enzymes and transcription factors, which are responsible for coordinating processes such as glycogen synthesis and cell adhesion. GSK-3β activity is also required for nuclear activity of Rel dimers, which mediate an anti-apoptotic response to TNFα in mice. GSK-3 catalytic kinase activity is controlled through differential phosphorylation of serine/threonine residues, which have an inhibitory effect, and tyrosine residues, which have an activating effect. Growth factor stimulation of mammalian cells expressing GSK-3α and GSK-3β induces phosphorylation of Ser 21 and Ser 9, respectively, through a phosphatidylinositol 3-kinase (PI 3-K)-protein kinase B (PKB)-dependent pathway, thereby enhancing proliferative signals. Additionally, GSK-3 physically associates with cAMP-dependent protein kinase A (PKA), which phosphorylates Ser 21 of GSK-3α or Ser 9 of GSK-3β and inactivates both forms. GSK-3α/β is positively regulated by phosphorylation on Tyr 279 and Tyr 216, respectively. Activated GSK-3α/β participates in energy metabolism, neuronal cell development, and body pattern formation. Tyrosine dephosphorylation of GSK-3 is involved in its extracellular signal-dependent inactivation.
Background References
1. Ji WT et al. Areca nut extract induces pyknotic necrosis in serum-starved oral cells via increasing reactive oxygen species and inhibiting GSK3 : an implication for cytopathic effects in betel quid chewers. PLoS One 8:e63295 (2013).
2. Yan X et al. Huaier aqueous extract inhibits ovarian cancer cell motility via the AKT/GSK3 / -catenin pathway. PLoS One 8:e63731 (2013).
Sequence Similarity
Belongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. GSK-3 subfamily.
Tissue Specificity
Expressed in testis, thymus, prostate and ovary and weakly expressed in lung, brain and kidney. Colocalizes with EIF2AK2/PKR and TAU in the Alzheimer disease (AD) brain.
Post-translational Modification
Phosphorylated by AKT1 and ILK1. Upon insulin-mediated signaling, the activated PKB/AKT1 protein kinase phosphorylates and desactivates GSK3B, resulting in the dephosphorylation and activation of GYS1. Activated by phosphorylation at Tyr-216. Inactivated by phosphorylation at Ser-9 (Probable). Phosphorylated in a circadian manner in the hippocampus (By similarity).; Mono-ADP-ribosylation by PARP10 negatively regulates kinase activity.
Subcellular Location
Cytoplasm, Nucleus, Cell membrane.
Synonyms
Glycogen Synthase Kinase 3 Beta antibody
Glycogen synthase kinase-3 beta antibody
GSK 3 beta antibody
GSK-3 beta antibody
GSK3B antibody
GSK3B_HUMAN antibody
GSK3beta isoform antibody
Serine/threonine-protein kinase GSK3B antibody
Images
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☑ Cell treatment (CT)
Western blot analysis of Phospho-GSK3 beta (S9) on different lysates with Rabbit anti-Phospho-GSK3 beta (S9) antibody (HA750128) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution.
Lane 1: HeLa cell lysate (15 µg/Lane)
Lane 2: HeLa treated with 100nM Calyculin A for 30 minutes cell lysate (15 µg/Lane)
Predicted band size: 47 kDa
Observed band size: 47 kDa
Exposure time: Lane 1-2 (left): 46 seconds; ECL: K1801;
Exposure time: Lane 1-2 (right): 45 seconds; ECL: K1802;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA750128) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
☑ Cell treatment (CT)
Western blot analysis of Phospho-GSK3 beta (S9) on different lysates with Rabbit anti-Phospho-GSK3 beta (S9) antibody (HA750128) at 1/5,000 dilution.
Lane 1: NIH/3T3 cell lysate (20 µg/Lane)
Lane 2: NIH/3T3 treated with 100ng/mL PDGF for 5 minutes cell lysate (20 µg/Lane)
Lane 3: C6 cell lysate (20 µg/Lane)
Lane 4: C6 treated with 100ng/mL PDGF for 5 minutes cell lysate (20 µg/Lane)
Lane 5: C6 cell lysate (20 µg/Lane)
Lane 6: C6 treated with 150nM insulin for 15 minutes cell lysate (20 µg/Lane)
Predicted band size: 47 kDa
Observed band size: 47 kDa
Exposure time: 6 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA750128) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
☑ Cell treatment (CT)
Immunocytochemistry analysis of HeLa cells treated with 100nM Calyculin A for 30 minutes labeling Phospho-GSK3 beta (S9) with Rabbit anti-Phospho-GSK3 beta (S9) antibody (HA750128) at 1/500 dilution.
Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Phospho-GSK3 beta (S9) antibody (HA750128) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
☑ Cell treatment (CT)
Immunocytochemistry analysis of NIH/3T3 cells treated with 100ng/mL PDGF for 5 minutes labeling Phospho-GSK3 beta (S9) with Rabbit anti-Phospho-GSK3 beta (S9) antibody (HA750128) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Phospho-GSK3 beta (S9) antibody (HA750128) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
☑ Cell treatment (CT)
Immunocytochemistry analysis of C6 cells treated with 100ng/mL PDGF for 5 minutes labeling Phospho-GSK3 beta (S9) with Rabbit anti-Phospho-GSK3 beta (S9) antibody (HA750128) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Phospho-GSK3 beta (S9) antibody (HA750128) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-Phospho-GSK3 beta (S9) antibody (HA750128) at 1/50 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750128) at 1/50 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-Phospho-GSK3 beta (S9) antibody (HA750128) at 1/50 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750128) at 1/50 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
☑ Cell treatment (CT)
Flow cytometric analysis of NIH/3T3 cells treated with 100ng/mL PDGF for 5 minutes labeling Phospho-GSK3 beta (S9).
Cells were fixed and permeabilized. Then stained with the primary antibody (HA750128, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
Citation
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Salvianolic Acid B Inhibited LH2 Expression to Reduce Collagen Synthesis in Pulmonary Fibrosis
Journal: Journal Of Cellular And Molecular Medicine
DOI: 10.1111/jcmm.71168
IF: 4.7
Application: WB
Reactivity: Human,Mouse
Publish date: 2026 May
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