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Phospho-GSK3B (Ser9) Recombinant Rabbit Monoclonal Antibody (3A6)

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Phospho-GSK3B (Ser9) Antibody in Immunocytochemistry (ICC/IF)

Phospho-GSK3B (Ser9) Antibody in Immunocytochemistry (ICC/IF)

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Phospho-GSK3B (Ser9) Antibody (MA5-33145) in ICC/IF

Immunofluorescence analysis of Glycogen synthase kinase-3 beta was performed using 70% confluent log phase HeLa and HeLa cells treated with Calyculin A (100 nm for 60 minutes). The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with Phospho-GSK3B (Ser9) Recombin... View More

Phospho-GSK3B (Ser9) Antibody in Immunocytochemistry (ICC/IF)

Phospho-GSK3B (Ser9) Antibody in Immunocytochemistry (ICC/IF)

Phospho-GSK3B (Ser9) Antibody in Immunohistochemistry (Paraffin) (IHC (P))

Phospho-GSK3B (Ser9) Antibody in Western Blot (WB)

Phospho-GSK3B (Ser9) Antibody in Western Blot (WB)

Phospho-GSK3B (Ser9) Antibody

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Phospho-GSK3B (Ser9) Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of Glycogen synthase kinase-3 beta was performed using 70% confluent log phase HeLa and HeLa cells treated with Calyculin A (100 nm for 60 minutes). The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with Phospho-GSK3B (Ser9) Recombinant Rabbit Monoclonal Antibody (Product # MA5-33145) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing Plasma membrane and Cytoplasmic localization. Panel e represents untreated HeLa cells with no signal. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Phospho-GSK3B (Ser9) Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescent analysis of Phospho-GSK3B (Ser9) in Hela cells (treated with 50mM Calyculin A for 30min) using a Phospho-GSK3B (Ser9) monoclonal antibody (Product # MA5-33145) at a dilution of 1:100. The cells were fixed in 4% formaldehyde, permeabilized using 0.2% Triton X-100 and blocked in 10% normal Goat Serum. The cells were then incubated with the antibody overnight at 4°C. The secondary antibody was Alexa Fluor 488-congugated Goat Anti-Rabbit IgG (H+L). Cells were counter-stained with DAPI.

Phospho-GSK3B (Ser9) Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemical analysis of Phospho-GSK3B (Ser9) in paraffin embedded human kidney tissue using a Phospho-GSK3B (Ser9) monoclonal antibody (Product # MA5-33145) at a dilution of 1:100. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4°C overnight. The primary is detected by a biotinylated secondary antibody and visualized using an HRP conjugated SP system.

Phospho-GSK3B (Ser9) Antibody in Western Blot (WB) — Western Blot analysis of Phospho-GSK3B (Ser9) using a Phospho-GSK3B (Ser9) Monoclonal antibody (Product # MA5-33145) at a concentration of 0.77 µg/mL. Positive WB detected in: Hela whole cell lysate,293 whole cell lysate,A549 whole cell lysate(treated with Calyculin A or not). All lanes: Phospho-GSK3B antibody at 0.77µg/ml. A secondary Goat polyclonal antibody to rabbit IgG was applied at a 1:50,000 dilution. Observed band size: 46 kDa.

Phospho-GSK3B (Ser9) Antibody in Western Blot (WB) — Western Blot was performed using Anti-Phospho-GSK3B (Ser9) Recombinant Rabbit Monoclonal Antibody (Product # MA5-33145) and a 50 kDa band corresponding to Glycogen synthase kinase-3 beta was observed to be induced upon Calyculin A treatment in HeLa cells, and upregulated upon EGF treatment and downregulated upon SB216763 followed by EGF treatment in A549 cells. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), HeLa treated with Calyculin A (100 nm for 60 minutes) (Lane 2), A549 (Lane 3), A549 treated with EGF (200 ng/mL for 30 minutes) (Lane 4) and A549 treated with SB216763 (10 µM for 30 minutes) followed by EGF (200 ng/mL for 30 mins) (Lane 5) were electrophoresed using NuPAGE™ 10% Bis-Tris Protein Gel (Product # NP0302BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23002) by iBlot® 2 Dry Blotting System (Product # IB21001). The Blot was probed with the primary antibody (1:3,500 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4,000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Phospho-GSK3B (Ser9) Antibody — Altered expression of proteins upon cell treatment demonstrates antibody specificity. Western blot using Phospho-GSK3B (Ser9) Recombinant Rabbit Monoclonal Antibody (Product # MA5-33145), shows induction of protein in HeLa cells upon Calyculin A treatment, and upregulation in A549 cells upon EGF treatment and downregulation upon SB216763 followed by EGF treatment. {TM}

Immunogen image

Phospho-GSK3B (Ser9) Recombinant Rabbit Monoclonal Antibody (3A6)

Immunogen Image Phospho-GSK3B (Ser9) Recombinant Rabbit Monoclonal Antibody (3A6)

产品信息

MA5-33145

应用

建议稀释比

已发表文章

免疫印迹 (WB)

1:500-1:5,000

-

免疫组化（石蜡） (IHC (P))

1:50-1:200

-

免疫细胞化学 (ICC/IF)

1:20-1:200

-

酶联免疫吸附实验 (ELISA)

Assay-Dependent

-

产品规格

种属反应

Human

宿主/亚型

Rabbit / IgG

Expression System

HEK293 cells

分类

Recombinant Monoclonal

类型

Antibody

克隆号

3A6

抗原

A synthesized peptide derived from human Phospho-GSK3B (Ser9)

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偶联物

Unconjugated Unconjugated Unconjugated

形式

Liquid

浓度

0.25 mg/mL

纯化类型

Affinity chromatography

保存液

PBS, pH 7.4, with 50% glycerol

内含物

0.02% sodium azide

保存条件

-20°C or -80°C if preferred

运输条件

Ambient (domestic); Wet ice (international)

靶标信息

GSK3B is a proline-directed serine/threonine protein kinase involved in energy metabolism, neuronal cell development and body pattern formation. It is a negative regulator of glucose homeostasis and is involved in energy metabolism, inflammation, ER-stress, mitochondrial dysfunction, and apoptotic pathways. GSK3B is a critical component of the Wnt/beta-catenin signaling pathway, and its activity results in the inhibition of Wnt signaling by mediating the degradation of beta-catenin. GSK3B is also involved in the Hedgehog, Notch, and insulin signaling pathways.

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引用该产品

生物信息学

蛋白别名: EC 2.7.11.26; Glycogen synthase kinase-3 beta; GSK 3 beta; GSK 3B; GSK-3 beta; GSK-3 beta GSK3B; GSK-3b; GSK3 beta; GSK3beta isoform; pp37; Serine/threonine-protein kinase GSK3B

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基因别名: GSK3B

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UniProt ID: (Human) P49841

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Entrez Gene ID: (Human) 2932

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RNA polymerase II transcription factor binding p53 binding protein kinase activity protein serine/threonine kinase activity integrin binding protein binding ATP binding beta-catenin binding kinase activity protein kinase binding ubiquitin protein ligase binding protein kinase A catalytic subunit binding ionotropic glutamate receptor binding tau protein binding tau-protein kinase activity NF-kappaB binding non-receptor serine/threonine protein kinase

re-entry into mitotic cell cycle epithelial to mesenchymal transition positive regulation of cell-matrix adhesion glycogen metabolic process regulation of gene expression by genetic imprinting protein phosphorylation protein export from nucleus ER overload response dopamine receptor signaling pathway axonogenesis myoblast fusion aging circadian rhythm organ morphogenesis negative regulation of cardiac muscle hypertrophy positive regulation of peptidyl-threonine phosphorylation positive regulation of mitochondrial membrane potential negative regulation of neuron maturation response to activity Wnt signaling pathway cell migration peptidyl-serine phosphorylation peptidyl-threonine phosphorylation hippocampus development establishment of cell polarity negative regulation of protein complex assembly positive regulation of protein complex assembly negative regulation of protein binding positive regulation of protein binding response to estradiol positive regulation of proteasomal ubiquitin-dependent protein catabolic process response to insulin regulation of microtubule-based process positive regulation of peptidyl-serine phosphorylation negative regulation of smooth muscle cell apoptotic process protein localization to microtubule intracellular signal transduction cellular response to hepatocyte growth factor stimulus cellular response to interleukin-3 cellular response to erythropoietin response to drug negative regulation of apoptotic process proteasome-mediated ubiquitin-dependent protein catabolic process negative regulation of MAP kinase activity positive regulation of neuron apoptotic process positive regulation of GTPase activity hypermethylation of CpG island canonical Wnt signaling pathway involved in positive regulation of apoptotic process fat cell differentiation negative regulation of glycogen biosynthetic process positive regulation of protein catabolic process positive regulation of axon extension positive regulation of transcription from RNA polymerase II promoter protein autophosphorylation positive regulation of protein export from nucleus regulation of neuronal synaptic plasticity positive regulation of smooth muscle cell proliferation negative regulation of dendrite morphogenesis negative regulation of nitric-oxide synthase activity negative regulation of NFAT protein import into nucleus canonical Wnt signaling pathway intrinsic apoptotic signaling pathway in response to endoplasmic reticulum stress cellular response to hydrogen peroxide superior temporal gyrus development cellular response to mechanical stimulus cellular response to iron(II) ion cellular response to lithium ion response to epinephrine negative regulation of canonical Wnt signaling pathway extrinsic apoptotic signaling pathway in absence of ligand chemical synaptic transmission, postsynaptic regulation of cellular response to heat negative regulation of protein localization to nucleus positive regulation of mitochondrial outer membrane permeabilization involved in apoptotic signaling pathway positive regulation of neuron death response to L-glutamate cellular response to dopamine negative regulation of dopaminergic neuron differentiation beta-catenin destruction complex assembly beta-catenin destruction complex disassembly response to insulin-like growth factor stimulus response to ultrasound response to angiotensin negative regulation of type B pancreatic cell development positive regulation of excitatory postsynaptic potential negative regulation of glycogen (starch) synthase activity positive regulation of DNA biosynthetic process positive regulation of stem cell differentiation negative regulation of neuron migration