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GSK3 beta Antibody (YA397)

2 Cited Publications
Customer Review

Based on 2 publication(s) in Google Scholar

GSK3 beta Antibody (YA397) is a Rabbit-derived and non-conjugated IgG monoclonal antibody, targeting to GSK3 beta.

For research use only. We do not sell to patients.
  • Host:

    Rabbit

  • Isotype:

    IgG

  • Application:

    WB, ICC/IF, IHC-P, FC

  • Reactivity :

    Human, Mouse, Rat

  • Formulation:

    Supplied in 1*TBS (pH7.4), 0.05% BSA and 40% Glycerol. Preservative: 0.05% Sodium Azide.

  • Conjugation:
    Non-conjugated
ALL WB IHC-P ICC FC
  • Experimental Validation Results for GSK3 beta Antibody (YA397)
  • Experimental Validation Results for GSK3 beta Antibody (YA397)
  • Experimental Validation Results for GSK3 beta Antibody (YA397)
  • Experimental Validation Results for GSK3 beta Antibody (YA397)
  • Experimental Validation Results for GSK3 beta Antibody (YA397)
  • Experimental Validation Results for GSK3 beta Antibody (YA397)
  • Experimental Validation Results for GSK3 beta Antibody (YA397)
  • GSK3 beta Antibody (YA397) Product Image
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Applications

Application
WB Info
WB: Western Blot
ICC/IF Info
ICC/IF: Immunocytochemistry/
Immunofluorescence
IHC-P Info
IHC-P: Immunohistochemistry-Paraffin
FC Info
FC: Flow Cytometry
Dilution Ratio 1:500-1:2000 1:50-1:200 1:50-1:200 1:50-1:100

Product Details

Description

GSK3 beta Antibody (YA397) is a Rabbit-derived and non-conjugated IgG monoclonal antibody, targeting to GSK3 beta.

  • Host Rabbit
  • Clonality Recombinant, Monoclonal
  • Species Reactivity
    Human, Mouse, Rat
  • Observed Molecular Weight
    Observed band size: 43 kDa Info
    Note: Due to possible protein modifications or aggregation, the molecular weight should be confirmed by actual measurement, and the predicted value is for reference only.
  • Calculated Molecular Weight Predicted band size: 47 kDa
Species Reactivity Database
Immunogen

Synthetic peptide corresponding to Human GSK3 beta.AA range:360-400.

Sensitivity

Endogenous

Purification

Protein A affinity purified.

Conjugation

Non-conjugated

Modification

Unmodified

Isotype

IgG

RRID

AB_3102570

Product Properties

  • Appearance

    Liquid

  • Formulation

    Supplied in 1*TBS (pH7.4), 0.05% BSA and 40% Glycerol. Preservative: 0.05% Sodium Azide.

  • Concentration

    Batch-dependent, Please check the COA for the concentration of each lot. Check Lot Concentration

  • Storage & Stability

    Stored at -20°C for 1 year. Avoid repeated freeze / thaw cycles.

  • Shipping

    Shipping with blue ice.

Verification Images

  • Experimental Validation Results for GSK3 beta Antibody (YA397)
    Western blot analysis was performed on protein extracts (25 μg) from HeLa (lane 2), NIH/3T3 (lane 3), 293T (lane 4), A549 (lane 5), Mouse brain (lane 6), and MCF-7 (lane 7) using GSK3 beta antibody. Proteins were transferred onto a 0.45 μm PVDF membrane using the Trans-Blot® Turbo™ system for 13 min. The membrane was then blocked with 5% nonfat milk in TBST (HY-K1025) for 1 h at room temperature. The primary antibody (1:1000) and loading control antibody GAPDH Antibody (HRP) (HY-P80954A) (1:5000) were diluted in 5% nonfat milk in TBST and incubated with the membrane overnight at 4°C. After washing, the membrane of primary antibody was incubated with HRP-conjugated goat anti-rabbit/mouse IgG secondary antibody (HY-P8001/HY-P8004) (1:5000) diluted in 5% nonfat milk in TBST for 1 h at room temperature. Protein bands were visualized using an Ultra High Sensitivity ECL detection kit (HY-K1005).
  • Experimental Validation Results for GSK3 beta Antibody (YA397)
    Western blot analysis of extracts from Hela (lane 2(20μg) ) , HEK-293 (lane 3(20μg) and C6 (lane 4(20μg) ) using GSK3 beta Rabbit mAb（HY-P80497）. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in TBST for 2 hour at room temperature. The primary antibody (1/1000) and Loading control antibody (Beta Actin , HY-P80438, 1/10000) was used in 5% BSA in TBST at 4°C overnight. Goat Anti-Rabbit IgG-HRP Secondary Antibody (1/10,000) was used for 1 hour at room temperature.
  • Experimental Validation Results for GSK3 beta Antibody (YA397)
    Immunohistochemical analysis of paraffin-embedded Mouse brain tissue using GSK3 beta Antibody (YA397). The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HY-P80497, 1/200) in 4℃ overnight. 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.
  • Experimental Validation Results for GSK3 beta Antibody (YA397)
    Immunohistochemical analysis of paraffin-embedded Mouse brain tissue using GSK3 beta Antibody (YA397). The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HY-P80497, 1/200) in 4℃ overnight. 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.
  • Experimental Validation Results for GSK3 beta Antibody (YA397)
    Immunocytochemistry analysis of DU145 cells labeling GSK3 beta with GSK3 beta antibody (HY-P80497) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with GSK3 beta antibody (HY-P80497) at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) 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. The Nuclear counterstain was DAPI (Blue).
  • Experimental Validation Results for GSK3 beta Antibody (YA397)
    Flow cytometric analysis of 1X10^6 Hela cells labeling GSK3 beta Antibody (HY-P80497, red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Then stained with the primary antibody at 1/50 dilution for an hour at 4℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L (HY-P8002) was used as the secondary antibody at 1/1,000 dilution for 30 minutes at 4℃. Rabbit IgG Isotype Control (HY-P80879, blue) was used as the isotype control, cells without incubation with primary antibody were used as the unlabeled control (black).
  • Experimental Validation Results for GSK3 beta Antibody (YA397)
    Immunocytochemistry analysis of Hela cells labeling GSK3 beta with GSK3 beta antibody (HY-P80497) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with GSK3 beta antibody (HY-P80497) at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) 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. The Nuclear counterstain was DAPI (Blue).

Background

  • Function

    GSK3 beta is a Constitutively active protein kinase that acts as a negative regulator in the hormonal control of glucose homeostasis, Wnt signaling and regulation of transcription factors and microtubules, by phosphorylating and inactivating glycogen synthase (GYS1 or GYS2), EIF2B, CTNNB1/beta-catenin, APC, AXIN1, DPYSL2/CRMP2, JUN, NFATC1/NFATC, MAPT/TAU and MACF1. Requires primed phosphorylation of the majority of its substrates. In skeletal muscle, contributes to insulin regulation of glycogen synthesis by phosphorylating and inhibiting GYS1 activity and hence glycogen synthesis. May also mediate the development of insulin resistance by regulating activation of transcription factors. Regulates protein synthesis by controlling the activity of initiation factor 2B (EIF2BE/EIF2B5) in the same manner as glycogen synthase. In Wnt signaling, GSK3B forms a multimeric complex with APC, AXIN1 and CTNNB1/beta-catenin and phosphorylates the N-terminus of CTNNB1 leading to its degradation mediated by ubiquitin/proteasomes. Phosphorylates JUN at sites proximal to its DNA-binding domain, thereby reducing its affinity for DNA. Phosphorylates NFATC1/NFATC on conserved serine residues promoting NFATC1/NFATC nuclear export, shutting off NFATC1/NFATC gene regulation, and thereby opposing the action of calcineurin. Phosphorylates MAPT/TAU on 'Thr-548', decreasing significantly MAPT/TAU ability to bind and stabilize microtubules. MAPT/TAU is the principal component of neurofibrillary tangles in Alzheimer disease. Plays an important role in ERBB2-dependent stabilization of microtubules at the cell cortex. Phosphorylates MACF1, inhibiting its binding to microtubules which is critical for its role in bulge stem cell migration and skin wound repair. Probably regulates NF-kappa-B (NFKB1) at the transcriptional level and is required for the NF-kappa-B-mediated anti-apoptotic response to TNF-alpha (TNF/TNFA). Negatively regulates replication in pancreatic beta-cells, resulting in apoptosis, loss of beta-cells and diabetes. Through phosphorylation of the anti-apoptotic protein MCL1, may control cell apoptosis in response to growth factors deprivation. Phosphorylates MUC1 in breast cancer cells, decreasing the interaction of MUC1 with CTNNB1/beta-catenin. Is necessary for the establishment of neuronal polarity and axon outgrowth. Phosphorylates MARK2, leading to inhibition of its activity. Phosphorylates SIK1 at 'Thr-182', leading to sustainment of its activity. Phosphorylates ZC3HAV1 which enhances its antiviral activity. Phosphorylates SNAI1, leading to its ubiquitination and proteasomal degradation. Phosphorylates SFPQ at 'Thr-687' upon T-cell activation. Phosphorylates NR1D1 st 'Ser-55' and 'Ser-59' and stabilizes it by protecting it from proteasomal degradation. Regulates the circadian clock via phosphorylation of the major clock components including BMAL1, CLOCK and PER2. Phosphorylates FBXL2 at 'Thr-404' and primes it for ubiquitination by the SCF(FBXO3) complex and proteasomal degradation. Phosphorylates CLOCK AT 'Ser-427' and targets it for proteasomal degradation. Phosphorylates BMAL1 at 'Ser-17' and 'Ser-21' and primes it for ubiquitination and proteasomal degradation. Phosphorylates OGT at 'Ser-3' or 'Ser-4' which positively regulates its activity. Phosphorylates MYCN in neuroblastoma cells which may promote its degradation. Regulates the circadian rhythmicity of hippocampal long-term potentiation and BMAL1 and PER2 expression. Acts as a regulator of autophagy by mediating phosphorylation of KAT5/TIP60 under starvation conditions, activating KAT5/TIP60 acetyltransferase activity and promoting acetylation of key autophagy regulators, such as ULK1 and RUBCNL/Pacer. Negatively regulates extrinsic apoptotic signaling pathway via death domain receptors. Promotes the formation of an anti-apoptotic complex, made of DDX3X, BRIC2 and GSK3B, at death receptors, including TNFRSF10B. The anti-apoptotic function is most effective with weak apoptotic signals and can be overcome by stronger stimulation. Phosphorylates E2F1, promoting the interaction between E2F1 and USP11, stabilizing E2F1 and promoting its activity. Phosphorylates mTORC2 complex component RICTOR at 'Ser-1235' in response to endoplasmic stress, inhibiting mTORC2. Phosphorylates mTORC2 complex component RICTOR at 'Thr-1695' which facilitates FBXW7-mediated ubiquitination and subsequent degradation of RICTOR. Phosphorylates FXR1, promoting FXR1 ubiquitination by the SCF(FBXO4) complex and FXR1 degradation by the proteasome. Phosphorylates interleukin-22 receptor subunit IL22RA1, preventing its proteasomal degradation[1][2][3][4][5][6][7][8][9][10][11][12][13][14][15][16][17][18][19][20][21][22][23][24][25][26].

  • Subcellular Localization

    Cytoplasm; Nucleus; Cell membrane

  • Expression


    Tissue_specificity:It is expressed in the testes, thymus, prostate, and ovaries, and expressed weakly in the lungs, brain, and kidneys. It co-localizes with EIF2AK2/PKR and TAU in the brain tissue of Alzheimer's disease (AD) .

  • Subunit

    Monomer. Interacts with ARRB2, DISC1 and ZBED3 (By similarity). Interacts with CABYR, MMP2, MUC1, NIN and PRUNE1. Interacts with AXIN1; the interaction mediates hyperphosphorylation of CTNNB1 leading to its ubiquitination and destruction. Interacts with and phosphorylates SNAI1. Interacts with DNM1L (via a C-terminal domain). Found in a complex composed of MACF1, APC, AXIN1, CTNNB1 and GSK3B (By similarity). Interacts with SGK3. Interacts with DAB2IP (via C2 domain); the interaction stimulates GSK3B kinase activation. Interacts (via C2 domain) with PPP2CA. Interacts with the CLOCK-BMAL1 heterodimer (PubMed:19946213). Interacts with the BMAL1 (PubMed:28903391). Interacts with CTNND2 (PubMed:19706605). Interacts with NCYM (PubMed:24391509). The complex composed, at least, of APC, CTNNB1 and GSK3B interacts with JPT1; the interaction requires the inactive form of GSK3B (phosphorylated at 'Ser-9') (PubMed:25169422). Forms a complex composed of PRKAR2A or PRKAR2B, GSK3B and GSKIP through GSKIP interaction; facilitates PKA-induced phosphorylation and regulates GSK3B activity (PubMed:20007971, PubMed:25920809, PubMed:27484798). Interacts with GSKIP (PubMed:16981698). Interacts with GID8 (PubMed:28829046). Interacts with PIWIL2 (By similarity). Interacts with LMBR1L (PubMed:31073040). Interacts with DDX3X (PubMed:18846110). Interacts with BIRC2 (PubMed:18846110). Interacts with TNFRSF10B; TNFRSF10B stimulation inhibits GSK3B kinase activity (PubMed:18846110). Interacts with RICTOR; the interaction results in phosphorylation of RICTOR at 'Thr-1695' by GSK3B which facilitates FBXW7-mediated ubiquitination and subsequent degradation of RICTOR (PubMed:25897075). Found in a complex with SLC39A6, SLC39A10 and with GSK3B that controls NCAM1 phosphorylation (By similarity). Interacts with PKP3 (via ARM repeats); the interaction may be involved in PKP3 protein degradation (PubMed:34058472)

  • SwissProt ID

    P49841

  • Gene ID
    2931 [NCBI]
  • Research Field

    Neuroscience

GSK3 beta Antibody (YA397) Related Classifications

MOQ
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100 mg

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Measured by its ability to inhibit anti-CD3-induced proliferation of stimulated CTLL-2 mouse cytotoxic T cells. The ED50 this effect is 0.8023 μg/ml, corresponding to a specific activity is 1246.42 units/mg.

Assay Types
  • 1/16

    MedChemExpress Validation
     Western blot analysis was performed on protein extracts (25 μg) from HeLa (lane 2), NIH/3T3 (lane 3), 293T (lane 4), A549 (lane 5), Mouse brain (lane 6), and MCF-7 (lane 7) using GSK3 beta antibody. Proteins were transferred onto a 0.45 μm PVDF membrane using the Trans-Blot® Turbo™ system for 13 min. The membrane was then blocked with 5% nonfat milk in TBST (HY-K1025) for 1 h at room temperature. The primary antibody (1:1000) and loading control antibody GAPDH Antibody (HRP) (HY-P80954A) (1:5000) were diluted in 5% nonfat milk in TBST and incubated with the membrane overnight at 4°C. After washing, the membrane of primary antibody was incubated with HRP-conjugated goat anti-rabbit/mouse IgG secondary antibody (HY-P8001/HY-P8004) (1:5000) diluted in 5% nonfat milk in TBST for 1 h at room temperature. Protein bands were visualized using an Ultra High Sensitivity ECL detection kit (HY-K1005).

  • 1/16

    MedChemExpress Validation
     Western blot analysis of extracts from Hela (lane 2(20μg) ) , HEK-293 (lane 3(20μg) and C6 (lane 4(20μg) ) using GSK3 beta Rabbit mAb（HY-P80497）. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in TBST for 2 hour at room temperature. The primary antibody (1/1000) and Loading control antibody (Beta Actin , HY-P80438, 1/10000) was used in 5% BSA in TBST at 4°C overnight. Goat Anti-Rabbit IgG-HRP Secondary Antibody (1/10,000) was used for 1 hour at room temperature.

  • 1/16

    MedChemExpress Validation
     Immunohistochemical analysis of paraffin-embedded Mouse brain tissue using GSK3 beta Antibody (YA397). The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HY-P80497, 1/200) in 4℃ overnight. 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.

  • 1/16

    MedChemExpress Validation
     Immunohistochemical analysis of paraffin-embedded Mouse brain tissue using GSK3 beta Antibody (YA397). The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HY-P80497, 1/200) in 4℃ overnight. 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.

  • 1/16

    MedChemExpress Validation
     Immunocytochemistry analysis of DU145 cells labeling GSK3 beta with GSK3 beta antibody (HY-P80497) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with GSK3 beta antibody (HY-P80497) at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) 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. The Nuclear counterstain was DAPI (Blue).

  • 1/16

    MedChemExpress Validation
     Flow cytometric analysis of 1X10^6 Hela cells labeling GSK3 beta Antibody (HY-P80497, red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Then stained with the primary antibody at 1/50 dilution for an hour at 4℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L (HY-P8002) was used as the secondary antibody at 1/1,000 dilution for 30 minutes at 4℃. Rabbit IgG Isotype Control (HY-P80879, blue) was used as the isotype control, cells without incubation with primary antibody were used as the unlabeled control (black).

  • 1/16

    MedChemExpress Validation
     Immunocytochemistry analysis of Hela cells labeling GSK3 beta with GSK3 beta antibody (HY-P80497) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with GSK3 beta antibody (HY-P80497) at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) 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. The Nuclear counterstain was DAPI (Blue).

  • GSK3 beta Antibody (YA397) Product Image
    1/16
  • Experimental Validation Results for GSK3 beta Antibody (YA397)
  • Experimental Validation Results for GSK3 beta Antibody (YA397)
  • Experimental Validation Results for GSK3 beta Antibody (YA397)
  • Experimental Validation Results for GSK3 beta Antibody (YA397)
  • Experimental Validation Results for GSK3 beta Antibody (YA397)
  • Experimental Validation Results for GSK3 beta Antibody (YA397)
  • Experimental Validation Results for GSK3 beta Antibody (YA397)
  • GSK3 beta Antibody (YA397) Product Image
  • Experimental Validation Results for GSK3 beta Antibody (YA397)
    1/1
    MedChemExpress Validation
     Western blot analysis was performed on protein extracts (25 μg) from HeLa (lane 2), NIH/3T3 (lane 3), 293T (lane 4), A549 (lane 5), Mouse brain (lane 6), and MCF-7 (lane 7) using GSK3 beta antibody. Proteins were transferred onto a 0.45 μm PVDF membrane using the Trans-Blot® Turbo™ system for 13 min. The membrane was then blocked with 5% nonfat milk in TBST (HY-K1025) for 1 h at room temperature. The primary antibody (1:1000) and loading control antibody GAPDH Antibody (HRP) (HY-P80954A) (1:5000) were diluted in 5% nonfat milk in TBST and incubated with the membrane overnight at 4°C. After washing, the membrane of primary antibody was incubated with HRP-conjugated goat anti-rabbit/mouse IgG secondary antibody (HY-P8001/HY-P8004) (1:5000) diluted in 5% nonfat milk in TBST for 1 h at room temperature. Protein bands were visualized using an Ultra High Sensitivity ECL detection kit (HY-K1005).
  • Experimental Validation Results for GSK3 beta Antibody (YA397)
    1/1
    MedChemExpress Validation
     Western blot analysis of extracts from Hela (lane 2(20μg) ) , HEK-293 (lane 3(20μg) and C6 (lane 4(20μg) ) using GSK3 beta Rabbit mAb（HY-P80497）. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in TBST for 2 hour at room temperature. The primary antibody (1/1000) and Loading control antibody (Beta Actin , HY-P80438, 1/10000) was used in 5% BSA in TBST at 4°C overnight. Goat Anti-Rabbit IgG-HRP Secondary Antibody (1/10,000) was used for 1 hour at room temperature.
  • Experimental Validation Results for GSK3 beta Antibody (YA397)
    1/1
    MedChemExpress Validation
     Immunohistochemical analysis of paraffin-embedded Mouse brain tissue using GSK3 beta Antibody (YA397). The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HY-P80497, 1/200) in 4℃ overnight. 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.
  • Experimental Validation Results for GSK3 beta Antibody (YA397)
    1/1
    MedChemExpress Validation
     Immunohistochemical analysis of paraffin-embedded Mouse brain tissue using GSK3 beta Antibody (YA397). The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HY-P80497, 1/200) in 4℃ overnight. 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.
  • Experimental Validation Results for GSK3 beta Antibody (YA397)
    1/1
    MedChemExpress Validation
     Immunocytochemistry analysis of DU145 cells labeling GSK3 beta with GSK3 beta antibody (HY-P80497) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with GSK3 beta antibody (HY-P80497) at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) 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. The Nuclear counterstain was DAPI (Blue).
  • Experimental Validation Results for GSK3 beta Antibody (YA397)
    1/1
    MedChemExpress Validation
     Flow cytometric analysis of 1X10^6 Hela cells labeling GSK3 beta Antibody (HY-P80497, red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Then stained with the primary antibody at 1/50 dilution for an hour at 4℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L (HY-P8002) was used as the secondary antibody at 1/1,000 dilution for 30 minutes at 4℃. Rabbit IgG Isotype Control (HY-P80879, blue) was used as the isotype control, cells without incubation with primary antibody were used as the unlabeled control (black).
  • Experimental Validation Results for GSK3 beta Antibody (YA397)
    1/1
    MedChemExpress Validation
     Immunocytochemistry analysis of Hela cells labeling GSK3 beta with GSK3 beta antibody (HY-P80497) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with GSK3 beta antibody (HY-P80497) at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) 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. The Nuclear counterstain was DAPI (Blue).
  • Experimental Validation Results for GSK3 beta Antibody (YA397)
  • Experimental Validation Results for GSK3 beta Antibody (YA397)
  • Experimental Validation Results for GSK3 beta Antibody (YA397)
  • Experimental Validation Results for GSK3 beta Antibody (YA397)
  • Experimental Validation Results for GSK3 beta Antibody (YA397)
  • Experimental Validation Results for GSK3 beta Antibody (YA397)
  • Experimental Validation Results for GSK3 beta Antibody (YA397)
  • GSK3 beta Antibody (YA397) Product Image
    MedChemExpress Validation

    Western blot analysis was performed on protein extracts (25 μg) from HeLa (lane 2), NIH/3T3 (lane 3), 293T (lane 4), A549 (lane 5), Mouse brain (lane 6), and MCF-7 (lane 7) using GSK3 beta antibody. Proteins were transferred onto a 0.45 μm PVDF membrane using the Trans-Blot® Turbo™ system for 13 min. The membrane was then blocked with 5% nonfat milk in TBST (HY-K1025) for 1 h at room temperature. The primary antibody (1:1000) and loading control antibody GAPDH Antibody (HRP) (HY-P80954A) (1:5000) were diluted in 5% nonfat milk in TBST and incubated with the membrane overnight at 4°C. After washing, the membrane of primary antibody was incubated with HRP-conjugated goat anti-rabbit/mouse IgG secondary antibody (HY-P8001/HY-P8004) (1:5000) diluted in 5% nonfat milk in TBST for 1 h at room temperature. Protein bands were visualized using an Ultra High Sensitivity ECL detection kit (HY-K1005).

  • GSK3 beta Antibody (YA397) Product Image
    MedChemExpress Validation

    Western blot analysis of extracts from Hela (lane 2(20μg) ) , HEK-293 (lane 3(20μg) and C6 (lane 4(20μg) ) using GSK3 beta Rabbit mAb（HY-P80497）. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in TBST for 2 hour at room temperature. The primary antibody (1/1000) and Loading control antibody (Beta Actin , HY-P80438, 1/10000) was used in 5% BSA in TBST at 4°C overnight. Goat Anti-Rabbit IgG-HRP Secondary Antibody (1/10,000) was used for 1 hour at room temperature.

  • GSK3 beta Antibody (YA397) Product Image
    MedChemExpress Validation

    Immunohistochemical analysis of paraffin-embedded Mouse brain tissue using GSK3 beta Antibody (YA397). The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HY-P80497, 1/200) in 4℃ overnight. 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.

  • GSK3 beta Antibody (YA397) Product Image
    MedChemExpress Validation

    Immunohistochemical analysis of paraffin-embedded Mouse brain tissue using GSK3 beta Antibody (YA397). The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HY-P80497, 1/200) in 4℃ overnight. 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.

  • GSK3 beta Antibody (YA397) Product Image
    MedChemExpress Validation

    Immunocytochemistry analysis of DU145 cells labeling GSK3 beta with GSK3 beta antibody (HY-P80497) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with GSK3 beta antibody (HY-P80497) at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) 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. The Nuclear counterstain was DAPI (Blue).

  • GSK3 beta Antibody (YA397) Product Image
    MedChemExpress Validation

    Flow cytometric analysis of 1X10^6 Hela cells labeling GSK3 beta Antibody (HY-P80497, red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Then stained with the primary antibody at 1/50 dilution for an hour at 4℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L (HY-P8002) was used as the secondary antibody at 1/1,000 dilution for 30 minutes at 4℃. Rabbit IgG Isotype Control (HY-P80879, blue) was used as the isotype control, cells without incubation with primary antibody were used as the unlabeled control (black).

  • GSK3 beta Antibody (YA397) Product Image
    MedChemExpress Validation

    Immunocytochemistry analysis of Hela cells labeling GSK3 beta with GSK3 beta antibody (HY-P80497) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with GSK3 beta antibody (HY-P80497) at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) 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. The Nuclear counterstain was DAPI (Blue).

  • GSK3 beta Antibody (YA397) Product Image WB
  • GSK3 beta Antibody (YA397) Product Image WB
  • GSK3 beta Antibody (YA397) Product Image IHC-P
  • GSK3 beta Antibody (YA397) Product Image IHC-P
  • GSK3 beta Antibody (YA397) Product Image ICC
  • GSK3 beta Antibody (YA397) Product Image FC
  • GSK3 beta Antibody (YA397) Product Image ICC