Phospho-GSK3 beta (S9) Recombinant Rabbit Monoclonal Antibody [SY02-71]
cat.: ET1607-60
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IF-Tissue, IHC-P
Clonality: Monoclonal
Clone number: SY02-71
Form: Liquid
Storage condition: 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.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 47 kDa
Isotype: IgG
Immunogen: Synthetic phospho-peptide corresponding to residues surrounding Ser9 of Human GSK3 beta aa 1-50 / 420.
Positive control: HeLa treated with 100nM Calyculin A for 30 minutes cell lysate, Hela, MCF-7, human colon carcinoma tissue, human breast tissue, human breast carcinoma tissue, human kidney tissue, human pancreas tissue.
Subcellular location: Cytoplasm, Nucleus, Cell membrane.
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
1:2,000-1:5,000
1:50-1:200
1:50-1:200
1:50-1:200
Uniprot #: SwissProt: P49841 Human | Q9WV60 Mouse | P18266 Rat
Alternative names: Glycogen Synthase Kinase 3 Beta Glycogen synthase kinase-3 beta GSK 3 beta GSK-3 beta GSK3B GSK3B_HUMAN GSK3beta isoform Serine/threonine-protein kinase GSK3B
Images
ET1607-60_1.jpg Fig1: Western blot analysis of Phospho-GSK3 beta (S9) on different lysates with Rabbit anti-Phospho-GSK3 beta (S9) antibody (ET1607-60) 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: 46 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 (ET1607-60) 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.
ET1607-60_2.jpg Fig2: Western blot analysis of Phospho-GSK3 beta(Ser 9) on Hela cell lysates.

Lane 1 : Hela cells, whole cell lysate, 10ug/lane
Lane 2 : Hela cells starved for 3 hours, whole cell lysates, 10ug/lane
Lane 3/4 : Hela cells starved for 3 hours, then treated with 100nM Calyculin A for 30 minutes, whole cell lysates, 10ug/lane
Lane 5 : Hela cells starved for 3 hours and treated with 100nM Calyculin A for 30 minutes, then treated with 2.8ug/ul lambda-PP for 30 minutes, whole cell lysates, 10ug/lane

All lanes :
Anti-Phospho-GSK3 beta(Ser 9) antibody (ET1607-60) at 1:500 dilution. Anti-GAPDH antibody (ET1601-4) at 1:10,000 dilution. Goat Anti-Rabbit IgG H&L (HRP) (HA1001) at 1/200,000 dilution.

Predicted band size: 47 kDa
Observed band size: 47 kDa

Blocking and diluting buffer: 5% BSA.

Exposure time: 1 minute 2 seconds
ET1607-60_3.jpg Fig3: ICC staining of Phospho-GSK3 beta (S9) in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1607-60, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1607-60_4.jpg Fig4: ICC staining of Phospho-GSK3 beta (S9) in MCF-7 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1607-60, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1607-60_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human colon carcinoma 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 (ET1607-60, 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.
ET1607-60_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human breast 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 (ET1607-60, 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.
ET1607-60_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded human breast carcinoma 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 (ET1607-60, 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.
ET1607-60_8.jpg Fig8: 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 (ET1607-60, 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.
ET1607-60_9.jpg Fig9: Immunohistochemical analysis of paraffin-embedded human pancreas 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 (ET1607-60, 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.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.