Anti-phospho-GSK3β Ser9 Antibody, clone 6K18 ZooMAb® Rabbit Monoclonal
SIGMA/ZRB1557 - recombinant, expressed in HEK 293 cells
Synonym: Glycogen synthase kinase-3 beta
Product Type: Chemical
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of untreated HeLa cells (A and B) and HeLa cells treated with phorbol 12-myristate 13-acetate (PMA)(C and D) cell lines were performed using a 1:100 dilution of Cat. No. ZRB1557, Anti-phospho-GSK3β Ser9, clone 6K18 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the cytoplasm.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of untreated HeLa cells (A and B) and HeLa cells treated with phorbol 12-myristate 13-acetate (PMA)(C and D) cell lines were performed using a 1:100 dilution of Cat. No. ZRB1557, Anti-phospho-GSK3β Ser9, clone 6K18 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the cytoplasm.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of untreated HeLa cells (A and B) and HeLa cells treated with phorbol 12-myristate 13-acetate (PMA)(C and D) cell lines were performed using a 1:100 dilution of Cat. No. ZRB1557, Anti-phospho-GSK3β Ser9, clone 6K18 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the cytoplasm.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of untreated HeLa cells (A and B) and HeLa cells treated with phorbol 12-myristate 13-acetate (PMA)(C and D) cell lines were performed using a 1:100 dilution of Cat. No. ZRB1557, Anti-phospho-GSK3β Ser9, clone 6K18 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the cytoplasm.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of untreated HeLa cells (A and B) and HeLa cells treated with phorbol 12-myristate 13-acetate (PMA)(C and D) cell lines were performed using a 1:100 dilution of Cat. No. ZRB1557, Anti-phospho-GSK3β Ser9, clone 6K18 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the cytoplasm.
Western Blotting Enhanced Validation - Recombinant Antibody Technology Lysates from untreated HeLa cells (Lane 1) and HeLa cells treated with phorbol 12-myristate 13-acetate (PMA; 500 nM; 30 min).(Lane 2) were probed with Cat. No. ZRB1557, Anti-phospho-GSK-3β Ser9, clone 6K18 ZooMAb® Rabbit Monoclonal (1:10,000 dilution). Proteins were visualized using a Donkey Anti-Rabbit IgG secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates phospho-GSK-3β-Ser9 (~45 kDa).
Western Blotting Enhanced Validation - Recombinant Antibody Technology Lysates from untreated NIH3T3 cells (Lane 1) and NIH3T3 cells treated with PDGF (Lane 2) were probed with Cat. No. ZRB1557, Anti-phospho-GSK-3β Ser9, clone 6K18 ZooMAb® Rabbit Monoclonal (1:10,000 dilution). Proteins were visualized using a Donkey Anti-Rabbit IgG secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates phospho-GSK-3β-Ser9 (~45 kDa).
Peptide Inhibition Assay Enhanced Validation - Recombinant Antibody Technology Lysates from HeLa cells treated with phorbol 12-myristate 13-acetate (PMA; 500 nM; 30 min) were probed using a 1:1,000 dilution of Cat. No. ZRB1557, Anti-phospho-GSK-3β Ser9, clone 6K18 ZooMAb® Rabbit Monoclonal. Proteins were visualized using a Donkey Anti-Rabbit IgG secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates phospho-GSK3β-Ser9 (~45 kDa). Lanes 1: Incubated without any peptide treatment. 2: Incubated with phospho-GSK-3β Ser9 peptide. 3: Incubated with unphosphorylated GSK-3b peptide.
12 Principles of Green Chemistry: Principle 1—Waste Prevention. This product has a smaller environmental footprint through reduction of waste in the manufacturing process.
12 Principles of Green Chemistry: Principle 4—Designing Safer Chemicals
Chemical products should be designed to affect their desired function while minimizing their toxicity.
Chemical products should be designed to affect their desired function while minimizing their toxicity.
12 Principles of Green Chemistry: Principle 6—Design for Energy Efficiency. This product was re-engineered to utilize less energy in the manufacturing process.
| antibody form | purified antibody |
| antibody product type | primary antibodies |
| biological source | rabbit |
| clone | 6K18, recombinant monoclonal |
| conjugate | unconjugated |
| description | recombinant, expressed in HEK 293 cells |
| enhanced validation | recombinant expression Learn more about Antibody Enhanced Validation  |
| epitope sequence | N-terminal |
| form | lyophilized |
| greener alternative category | Aligned |
| greener alternative product characteristics | Waste Prevention Designing Safer Chemicals Design for Energy Efficiency Learn more about the Principles of Green Chemistry . |
| isotype | IgG |
| mol wt | calculated mol wt 46.74 kDa |
| observed mol wt ~45 kDa | |
| packaging | antibody small pack of 25 μL |
| product line | ZooMAb® learn more |
| Protein ID accession no. | NP_002084  |
| purified by | using Protein A |
| Quality Level | 200 |
| recombinant | expressed in HEK 293 cells |
| shipped in | ambient |
| species reactivity | human, mouse |
| species reactivity (predicted by homology) | rat |
| storage temp. | 2-8°C |
| technique(s) | immunocytochemistry: suitable |
| peptide synthesis: suitable | |
| western blot: suitable | |
| UniProt accession no. | P49841 |
| Application: | Quality Control Testing Evaluated by Western Blotting in lysate from HeLa cells treated with phorbol 12-myristate 13-acetate (PMA). Western Blotting Analysis (WB): A 1:10,000 dilution of this antibody detected GSK-3β phosphorylated on serine 9 in lysate from HeLa cells treated with phorbol 12-myristate 13-acetate (PMA; 500 nM; 30 min). Tested applications Western Blotting Analysis: A 1:10,000 dilution from a representative lot detected phospho-GSK3β Ser9 in lysate from NIH3T3 cells treated with PDGF. Immunocytochemistry Analysis: A 1:100 dilution from a representative lot detected phospho-GSK3β-Ser9 in HeLa cells treated with phorbol 12-myristate 13-acetate (PMA).. Peptide Inhibition Assay: Target band detection in lysate from HeLa cell treated with phorbol 12-myristate 13-acetate m(PMA) was prevented by preblocking of a representative lot with the immunogen phosphopeptide, but not the corresponding non-phosphopeptide. Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user |
| Disclaimer: | Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals. |
| General description: | We are committed to bringing you greener alternative products, which adhere to one or more of The 12 Principles of Green Chemistry.This antibody is Preservative-free, produced without the harm or sacrifice of animals and exceptionally stable to allow for ambient shipping and storage if needed and thus aligns with "Waste Prevention", "Designing Safer Chemicals" and "Design for Energy Efficiency". Click here for more information. |
| General description: | ZooMAb® antibodies represent an entirely new generation of recombinant monoclonal antibodies. Each ZooMAb® antibody is manufactured using our proprietary recombinant expression system, purified to homogeneity, and precisely dispensed to produce robust and highly reproducible lot-to-lot consistency. Only top-performing clones are released for use by researchers. Each antibody is validated for high specificity and affinity across multiple applications, including its most commonly used application. ZooMAb® antibodies are reliably available and ready to ship when you need them. |
| Immunogen: | KLH-conjugated linear peptide corresponding to 9 amino acids surrounding phosphoserine 9 in human GSK-3b. |
| Legal Information: | ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany |
| Physical form: | Purified recombinant rabbit monoclonal antibody IgG, lyophilized in PBS, 5% Trehalose, normal appearance a coarse or translucent resin. The PBS/trehalose components in the ZooMAb formulation can have the appearance of a semi-solid (bead like gel) after lyophilization. This is a normal phenomenon. Please follow the recommended reconstitution procedure in the data sheet to dissolve the semi-solid, bead-like, gel-appearing material. The resulting antibody solution is completely stable and functional as proven by full functional testing. Contains no biocide or preservatives, such as azide, or any animal by-products. Larger pack sizes provided as multiples of 25 μL. |
| Reconstitution: | 300 μg/mL after reconstitution at 25 μL per vial. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type. |
| Specificity: | Clone 6K18 is a ZooMAb® Rabbit recombinant monoclonal antibody that specifically detects GSK-3b phosphorylated on serine 9. |
| Storage and Stability: | Recommend storage of lyophilized product at 2-8°C; Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial; Reconstitute each vial by adding 25 μL of filtered lab grade water or PBS; Reconstituted antibodies can be stored at 2-8°C, or -20°C for long term storage. Avoid repeated freeze-thaws. |
| Target description: | Glycogen synthase kinase-3 beta (UniProt: P49841; also known as EC:2.7.11.26, GSK-3 beta, Serine/threonine-protein kinase GSK3B) is encoded by the GSK3B gene (Gene ID: 2932) in human. GSK-3 is a multifunctional serine/threonine kinase, is a key regulator of numerous signaling pathways during embryogenesis and in metabolic control. Two isoforms of GSK-3 are reported in mammals: a 51 kDa GSK-3a and a 47 kDa GSK-3b. These two isoforms exhibit about 98% homology in their kinase domains but share only about 36% identity in the last 76 C-terminal amino acid residues. GSK-3a contains a glycine-rich extension at its N-terminal. GSK-3 is constitutively active in cells and is regulated through inhibition of its activity. It shows a preference for target proteins that are pre-phosphorylated at a priming residue located C-terminal to the site of GSK-3 phosphorylation. Priming phosphorylation, although not absolutely required, enhances the efficiency of phosphorylation of most GSK-3 substrates. Phosphorylation of a threonine residue in the activation loop (T-loop) is considered to be essential for its activity. The T-loop of GSK-3a is phosphorylated at Tyr279 and GSK-3b at Tyr216, which play a role in forcing open the substrate-binding site of the enzyme. A primary mechanism of GSK3 activity regulation is phosphorylation of N-terminal serine residues (Serine 21 in GSK-3a and serine 9 in GSK-3b). Phosphorylation of these sites is inhibitory to GSK3 kinase activity because the phosphorylated N-terminus acts as a competitive inhibitor for primed substrates. GSK-3b is constitutively active in resting cells and treatment of cells with an agent, such as insulin, causes GSK-3 inactivation through a PI 3-kinase (PI 3-K)-dependent mechanism. This ZooMAb® recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. |
| Storage Temp. | 2-8°C |
| UNSPSC | 12352203 |