WB
Western blot analysis of Phospho-GSK3 beta (S9) on different lysates with Rabbit anti-Phospho-GSK3 beta (S9) antibody at 1/1,000 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) Exposure time: 6 seconds; 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 at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-Phospho-GSK3 beta (S9) antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (1/50) for 30 minutes 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.ICC/IF
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 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 t 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.FC
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 (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 at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).| Product Name | Phospho-GSK3 beta (S9) Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Synthetic phospho-peptide corresponding to residues surrounding Ser9 of Human GSK3 beta aa 1-50 / 420. |
| Modification | p-S9 |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | FCICC/IFIHCWB |
| WB:1:1000-1:5000 IHC:1:50-1:200 ICC:1:50-1:200 FC:1:1000 |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | GSK3B |
|---|---|
| Gene Full Name | glycogen synthase kinase 3 beta |
| Gene Summary | The protein encoded by this gene is a serine-threonine kinase belonging to the glycogen synthase kinase subfamily. It is a negative regulator of glucose homeostasis and is involved in energy metabolism, inflammation, ER-stress, mitochondrial dysfunction, and apoptotic pathways. Defects in this gene have been associated with Parkinson disease and Alzheimer disease. [provided by RefSeq, Aug 2017] |
| Alternative Names | 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 |
| Molecular Weight(MW) | 47kDa |
| Cellular Localization | Cytoplasm, Nucleus, Cell membrane. |
WB
Western blot analysis of Phospho-GSK3 beta (S9) on different lysates with Rabbit anti-Phospho-GSK3 beta (S9) antibody at 1/1,000 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) Exposure time: 6 seconds; 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 at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-Phospho-GSK3 beta (S9) antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (1/50) for 30 minutes 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.
ICC/IF
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 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 t 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.
FC
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 (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 at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).| Application Notes | WB:1:1000-1:5000 IHC:1:50-1:200 ICC:1:50-1:200 FC:1:1000 |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
