Phospho-GSK3 alpha (S21) Recombinant Rabbit Monoclonal Antibody [PSH09-11]
cat.: HA723069
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse |
| Applications: | WB, IHC-P, Dot Blot |
| Clonality: | Monoclonal |
| Clone number: | PSH09-11 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage buffer: | PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 51 kDa |
| Isotype: | IgG |
| Immunogen: | Synthetic phospho-peptide corresponding to residues surrounding Ser21 of Human GSK3 alpha. |
| Positive control: | A549 treated with 100nM Calyculin A for 30 minutes cell lysate, SH-SY5Y treated with 200nM Calyculin A and 1μM Okadaic Acid for 1 hour cell lysate, NIH/3T3 starved overnight cell lysate, NIH/3T3 starved overnight then treated with 100ng/mL PDGF for 5 minutes cell lysate, mouse testis tissue, human breast tissue, human colon cancer tissue. |
| Subcellular location: | Apical dendrite, axon, cytoplasm, cytosol, mitochondrion, neuronal cell body, nucleus, postsynapse, proximal dendrite. |
| Recommended Dilutions:
WB IHC-P Dot Blot |
1:2,000 1:200-1:500 1:1,000 |
| Uniprot #: | SwissProt: P49840 Human | Q2NL51 Mouse |
| Alternative names: | DKFZp686D0638 Glycogen synthase kinase 3 alpha Glycogen synthase kinase-3 alpha GSK 3 alpha GSK 3A GSK-3 alpha Gsk3a GSK3A_HUMAN GSK3alpha Serine/threonine protein kinase GSK3A |
Images
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Fig1:
Western blot analysis of Phospho-GSK3 alpha (S21) on different lysates with Rabbit anti-Phospho-GSK3 alpha (S21) antibody (HA723069) at 1/2,000 dilution. Lane 1: A549 cell lysate Lane 2: A549 treated with 100nM Calyculin A for 30 minutes cell lysate Lane 3: SH-SY5Y cell lysate Lane 4: SH-SY5Y treated with 200nM Calyculin A and 1μM Okadaic Acid for 1 hour cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 51 kDa Observed band size: 51 kDa Exposure time: 1 minute; ECL: K1801; 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 (HA723069) at 1/2,000 dilution was 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. |
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Fig2:
Western blot analysis of Phospho-GSK3 alpha (S21) on different lysates with Rabbit anti-Phospho-GSK3 alpha (S21) antibody (HA723069) at 1/2,000 dilution. Lane 1: NIH/3T3 starved overnight cell lysate Lane 2: NIH/3T3 starved overnight then treated with 100ng/mL PDGF for 5 minutes cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 51 kDa Observed band size: 51 kDa Exposure time: 3 minutes; ECL: K1801; 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 (HA723069) at 1/2,000 dilution was 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. |
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Fig3:
Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-Phospho-GSK3 alpha (S21) antibody (HA723069) at 1/500 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723069) at 1/500 dilution for 1 hour 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. |
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Fig4:
Immunohistochemical analysis of paraffin-embedded human breast tissue with Rabbit anti-Phospho-GSK3 alpha (S21) antibody (HA723069) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723069) at 1/200 dilution for 1 hour 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. |
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Fig5:
Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Rabbit anti-Phospho-GSK3 alpha (S21) antibody (HA723069) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723069) at 1/200 dilution for 1 hour 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. |
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Fig6:
Dot blot analysis of Phospho-GSK3 alpha (S21) on different peptides with Rabbit anti-Phospho-GSK3 alpha (S21) antibody (HA723069) at 1/1,000 dilution. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution for 1 hour at room temperature. Lane 1: Phospho-GSK3 alpha (S21) peptide (positive) Lane 2: Unmodified GSK3 alpha peptide (negative) Lane 3: Phospho-GSK3 beta (S9) peptide (negative) Lane 4: Unmodified GSK3 beta peptide (negative) Proteins loading: 100ng, 25ng, 5ng; Blocking and dilution buffer: 5% NFDM/TBST; Exposure time: 10 seconds; ECL: K1801. |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.