PRODUCT CODE: ET1607-60

Phospho-GSK3 beta (Ser 9) Recombinant Rabbit Monoclonal Antibody [SY02-71] (ET1607-60)

  • Recombinant

Applications

  • WB

  • ICC/IF

  • IHC-P

REACTIVITY

  • Human

Western blot analysis of Phospho-GSK3 beta(Ser 9) on Hela cell lysates.<br />
<br />
Lane 1 : Hela cells, whole cell lysate, 10ug/lane<br />
Lane 2 : Hela cells starved for 3 hours, whole cell lysates, 10ug/lane<br />
Lane 3/4 : Hela cells starved for 3 hours, then treated with 100nM Calyculin A for 30 minutes, whole cell lysates, 10ug/lane<br />
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<br />
<br />
All lanes :<br />
Anti-Phospho-GSK3 beta(Ser 9) antibody (ET1603-40) 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.<br />
<br />
Predicted band size: 47 kDa<br />
Observed band size: 47 kDa<br />
<br />
Blocking and diluting buffer: 5% BSA.<br />
<br />
Exposure time: 1 minute 2 seconds
  • Western blot analysis of Phospho-GSK3 beta(Ser 9) on Hela cell lysates.<br />
<br />
Lane 1 : Hela cells, whole cell lysate, 10ug/lane<br />
Lane 2 : Hela cells starved for 3 hours, whole cell lysates, 10ug/lane<br />
Lane 3/4 : Hela cells starved for 3 hours, then treated with 100nM Calyculin A for 30 minutes, whole cell lysates, 10ug/lane<br />
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<br />
<br />
All lanes :<br />
Anti-Phospho-GSK3 beta(Ser 9) antibody (ET1603-40) 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.<br />
<br />
Predicted band size: 47 kDa<br />
Observed band size: 47 kDa<br />
<br />
Blocking and diluting buffer: 5% BSA.<br />
<br />
Exposure time: 1 minute 2 seconds
  • Western blot analysis of Phospho-GSK3 beta(Ser 9) on different lysates with Rabbit anti-Phospho-GSK3 beta(Ser 9) antibody (ET1607-60) at 1/500 dilution.<br />
<br />
Lane 1: Hela cell lysate<br />
Lane 2: A549 cell lysate<br />
<br />
Lysates/proteins at 10 µg/Lane.<br />
<br />
Predicted band size: 47 kDa<br />
Observed band size: 47 kDa<br />
<br />
Exposure time: 2 minutes;<br />
<br />
10% SDS-PAGE gel.<br />
<br />
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 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.
  • ICC staining of Phospho-GSK3 beta(Ser 9) 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).
  • ICC staining of Phospho-GSK3 beta(Ser 9) 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).
  • Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue using anti-Phospho-GSK3 beta(Ser 9) 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.
  • Immunohistochemical analysis of paraffin-embedded human breast tissue using anti-Phospho-GSK3 beta(Ser 9) 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.
  • Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using anti-Phospho-GSK3 beta(Ser 9) 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.
  • Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-Phospho-GSK3 beta(Ser 9) 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.
  • Immunohistochemical analysis of paraffin-embedded human pancreas tissue using anti-Phospho-GSK3 beta(Ser 9) 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.
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 (ET1603-40) 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

Applications

  • WB

  • ICC/IF

  • IHC-P

REACTIVITY

  • Human

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

Phospho-GSK3 beta (Ser 9) Recombinant Rabbit Monoclonal Antibody [SY02-71] (ET1607-60)

Immunogen

Synthetic phospho-peptide corresponding to residues surrounding ser9 of human gsk3 beta aa 1-50 / 420.

Host

Rabbit

Modification

Phospho

Modification Site

Ser 9

Positive Control

Hela cell lysate, A549 cell lysate, Hela, MCF-7, human colon carcinoma tissue, human breast tissue, human breast carcinoma tissue, human kidney tissue, human pancreas tissue.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

SY02-71

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C or -80C. 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

APPLICATION DILUTION

  • WB

  • 1:500-1:2,000

  • ICC/IF

  • 1:50-1:200

  • IHC-P

  • 1:50-1:200

TARGET

UNIPROT #

PROTEIN NAME

Phospho-GSK3 beta (Ser 9)

SYNONYMS

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

SEQUENCE SIMILARITIES

Belongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. GSK-3 subfamily.

TISSUE SPECIFICITY

Expressed in testis, thymus, prostate and ovary and weakly expressed in lung, brain and kidney. Colocalizes with EIF2AK2/PKR and TAU in the Alzheimer disease (AD) brain.

POST-TRANSLATIONAL MODIFICATION

Phosphorylated by AKT1 and ILK1. Upon insulin-mediated signaling, the activated PKB/AKT1 protein kinase phosphorylates and desactivates GSK3B, resulting in the dephosphorylation and activation of GYS1. Activated by phosphorylation at Tyr-216. Inactivated by phosphorylation at Ser-9 (Probable). Phosphorylated in a circadian manner in the hippocampus (By similarity).; Mono-ADP-ribosylation by PARP10 negatively regulates kinase activity.

SUBCELLULAR LOCATION

Cytoplasm, Nucleus, Cell membrane.

FUNCTION

Glycogen synthase kinase-3α (GSK-3α) and GSK-3β are highly similar isoforms of serine/ threonine kinases that regulate metabolic enzymes and transcription factors, which are responsible for coordinating processes such as glycogen synthesis and cell adhesion. GSK-3β activity is also required for nuclear activity of Rel dimers, which mediate an anti-apoptotic response to TNFα in mice. GSK-3 catalytic kinase activity is controlled through differential phosphorylation of serine/threonine residues, which have an inhibitory effect, and tyrosine residues, which have an activating effect. Growth factor stimulation of mammalian cells expressing GSK-3α and GSK-3β induces phosphorylation of Ser 21 and Ser 9, respectively, through a phosphatidylinositol 3-kinase (PI 3-K)-protein kinase B (PKB)-dependent pathway, thereby enhancing proliferative signals. Additionally, GSK-3 physically associates with cAMP-dependent protein kinase A (PKA), which phosphorylates Ser 21 of GSK-3α or Ser 9 of GSK-3β and inactivates both forms. GSK-3α/β is positively regulated by phosphorylation on Tyr 279 and Tyr 216, respectively. Activated GSK-3α/β participates in energy metabolism, neuronal cell development, and body pattern formation. Tyrosine dephosphorylation of GSK-3 is involved in its extracellular signal-dependent inactivation.

CITATIONS

  • Guodong Chen;Longquan Shao

    Rapamycin-Induced Autophagy Promotes the Chondrogenic Differentiation of Synovium-Derived Mesenchymal Stem Cells in the Temporomandibular Joint in Response to IL-1β

  • Xue, F., Zhao, Z., Gu, Y., ...

    7,8-Dihydroxyflavone modulates bone formation and resorption and ameliorates ovariectomy-induced osteoporosis. eLife, 10, e64872.

  • Jian Li; Caiqiao Zhang

    Metformin Prevents Follicular Atresia in Aging Laying Chickens through Activation of PI3K/AKT and Calcium Signaling Pathways