Invitrogen

GSK3B Recombinant Polyclonal Antibody (1HCLC)

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This Antibody was verified by KnockDown to ensure that the antibody binds to the antigen stated. View Details

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GSK3B Antibody in Immunofluorescence (IF) — Immunofluorescence analysis of GSK3B was performed using 70% confluent log phase A-431 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with GSK3 Rabbit Polyclonal Antibody (Product # 710132) at 5 µg/mL in 0.1% BSA and incubated overnight at 4 degree and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

GSK3B Antibody in Western Blot (WB) — Western blot analysis of GSK-3-beta was performed by loading 30 µg of HeLa, HEK-293, Jurkat, MCF-7, DU 145, A431, COS-7, and COLO 205 cell lysates using Novex®NuPAGE®4-12% Bis-Tris gel (Product # NP0321BOX), XCell SureLock Electrophoresis System (Product # EI0002), Novex® Sharp Pre-Stained Protein Standard (Product # LC5800), and iBlot® Dry Blotting System (Product # IB21001). Proteins were transferred to a nitrocellulose membrane and blocked with 5% skim milk for 1 hour at room temperature. GSK-3-beta was detected at ~47 kDa using GSK-3-beta Recombinant Rabbit Polyclonal Antibody (Product # 710132) at a 1:1000 dilution in 2.5% skim milk at 4°C overnight on a rocking platform. Detection was performed using an HRP-conjugated Goat anti-Rabbit secondary antibody (Product # G-21234) at a 1:5000 dilution and chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).

GSK3B Antibody in Western Blot (WB) — Knockdown of GSK3B was achieved by transfecting HeLa cells with GSK3B specific siRNAs (Silencer® select Product # s6240). Western blot analysis (Fig a) was performed using membrane enriched extracts from the GSK3B knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with Anti-GSK3B Recombinant Rabbit Polyclonal Antibody (Product # 710132, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25µg/mL, 1:4000 dilution). Densitometric analysis of this Western blot is shown in histogram (Fig b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to GSK3B.

GSK3B Antibody in Western Blot (WB) — Western blot analysis was performed on whole cell extracts of MCF7 (Lane 1), MDA-MB-231 (Lane 2), HeLa (Lane 3), Neuro-2a (Lane 4), U-87 MG (Lane 5), A-431 (Lane 6) and NIH/3T3 (Lane 7). The blot was probed with Anti-GSK3 antibody (Product # 710132, 1µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). A 47 kDa band corresponding to GSK3B was observed across the cell lines tested.

GSK3B Antibody in Western Blot (WB) — Western blot analysis of GSK3 in whole cell extracts of HepG2 (lane 1) and NIH-3T3 (lane 2) using a GSK3 Recombinant Rabbit Polyclonal Antibody (Product # 710132) at a dilution of 1 µg/mL. Samples were detected using chemiluminescence (ECL). Results show a band at ~53kDa.

GSK3B Antibody in KnockDown — Antibody specificity was demonstrated by siRNA mediated knockdown of target protein. HeLa cells were transfected with GSK3B siRNA and loss of signal was observed in Western Blot using Anti-GSK3B Oligo clonal Antibody (Product # 710132). {KD}

Product Details

Tested Applications

Dilution

Immunocytochemistry (ICC)

5 µg/mL

Immunofluorescence (IF)

5 µg/mL

Western Blot (WB)

1 µg/mL

Product Specifications

Species Reactivity

Human, Mouse, Non-human primate

Host / Isotype

Rabbit / IgG

Class

Recombinant Polyclonal

Type

Antibody

Clone

1HCLC

Immunogen

Recombinant protein corresponding to amino acids 317–433 of human GSK3 beta

Conjugate

Unconjugated

Form

Liquid

Concentration

0.5 mg/mL

Purification

Protein A

Storage buffer

PBS

Contains

0.09% sodium azide

Storage conditions

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

RRID

AB_2532582

Product Specific Information

This antibody is predicted to react with equine, mouse, rabbit and rat based on sequence homology.

Intact IgG appears on a non-reducing gel as ~150 kDa band and upon reduction generating a ~25 kDa light chain band and a ~50 kDa heavy chain.

Recombinant rabbit polyclonal antibodies are unique offerings from Thermo Fisher Scientific. They are comprised of a selection of multiple different recombinant monoclonal antibodies, providing the best of both worlds – the sensitivity of polyclonal antibodies with the specificity of monoclonal antibodies - all delivered with the consistency only found in a recombinant antibody. While functionally the same as a polyclonal antibody – recognizing multiple epitope sites on the target and producing higher detection sensitivity for low abundance targets – a recombinant rabbit polyclonal antibody has a known mixture of light and heavy chains. The exact population can be produced in every lot, circumventing the biological variability typically associated with polyclonal antibody production.

Target Information

Glycogen synthase kinase-3 beta (GSK3 beta) belongs to the subfamily of glycogen synthase kinases, that play a central role in energy metabolism, neuronal cell development, and body pattern formation. GSK3 has alpha and beta isoforms (51 kDa and 47 kDa respectively), that are functionally dissimilar. GSK3 beta plays a role in insulin and Wnt signaling, and is inhibited following the extracellular signaling events.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Bioinformatics

Protein Aliases: EC 2.7.11.26; Glycogen Synthase Kinase 3 beta; Glycogen synthase kinase-3 beta; GSK-3 beta; GSK-3 beta Factor A; GSK3B; GSK3beta isoform; Serine/threonine-protein kinase GSK3B

Gene Aliases: 7330414F15Rik; 8430431H08Rik; C86142; GSK-3; GSK-3beta; GSK3; GSK3B

UniProt ID: (Human) P49841, (Mouse) Q9WV60

Entrez Gene ID: (Human) 2932, (Mouse) 56637

Molecular Function: kinase non-receptor serine/threonine protein kinase protein kinase transferase

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 carbohydrate metabolic process cytoskeleton organization establishment or maintenance of cell polarity multicellular organism development nervous system development cell proliferation negative regulation of signal transduction regulation of neuron projection development negative regulation of neuron projection development myotube differentiation phosphorylation cell differentiation positive regulation of apoptotic process rhythmic process

Process(es)

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 nucleotide binding transcription factor binding transferase activity transferase activity, transferring phosphorus-containing groups