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Phospho-GSK3 alpha (S21) Recombinant Rabbit Monoclonal Antibody [PSH09-11]

Usd: 385

Specification

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Catalog# HA723069

Phospho-GSK3 alpha (S21) Recombinant Rabbit Monoclonal Antibody [PSH09-11]

Application

  • WB

  • IHC-P

  • Dot Blot

Reactivity

  • Human

  • Mouse

BSA and Azide free
Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

Phospho-GSK3 alpha (S21) Recombinant Rabbit Monoclonal Antibody [PSH09-11]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic phospho-peptide corresponding to residues surrounding Ser21 of Human GSK3 alpha.

Species Reactivity

Human, Mouse

Validated Applications

WB, IHC-P, Dot Blot

Molecular Weight

Predicted band size: 51 kDa

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.

Conjugation

unconjugated

Clone Number

PSH09-11

Reactivity Data

WB IHC-P Dot Blot
Human
Tested Tested
Tested Tested
Mouse
Tested Tested
Tested Tested

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:2,000

  • IHC-P

  • 1:200-1:500

  • Dot Blot

  • 1:1,000

Target

Function

Glycogen synthase kinase-3 alpha is an enzyme that in humans is encoded by the GSK3A gene. Glycogen synthase kinase 3-alpha EC 2.7.1.37 is a multifunctional protein serine kinase, homologous to Drosophila 'shaggy' (zeste-white3) and implicated in the control of several regulatory proteins including glycogen synthase and various transcription factors (e.g., JUN). It also plays a role in the WNT and phosphoinositide 3-kinase (especially PIK3CG) signaling pathways.

Background References

1. Wang J et al. Quantitative phosphoproteomics reveals GSK3A substrate network is involved in the cryodamage of sperm motility. Biosci Rep. 2021 Oct

2. Zhang W et al. Active Ingredient Paeonol of Jijiu Huiyang Decoction Alleviates Isoproterenol-Induced Chronic Heart Failure via the GSK3A/PPARalpha Pathway. Oxid Med Cell Longev. 2023 Feb

Subcellular Location

Apical dendrite, axon, cytoplasm, cytosol, mitochondrion, neuronal cell body, nucleus, postsynapse, proximal dendrite.

UNIPROT

SwissProt: P49840 Human

SwissProt: Q2NL51 Mouse

Synonyms

DKFZp686D0638 antibody

Glycogen synthase kinase 3 alpha antibody

Glycogen synthase kinase-3 alpha antibody

GSK 3 alpha antibody

GSK 3A antibody

GSK-3 alpha antibody

Gsk3a antibody

GSK3A_HUMAN antibody

GSK3alpha antibody

Serine/threonine protein kinase GSK3A antibody

Images

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of Phospho-GSK3 alpha (S21) on different lysates with Rabbit anti-Phospho-GSK3 alpha (S21) antibody (<a href="/products/HA723069" style="font-weight: bold;text-decoration: underline;">HA723069</a>) at 1/2,000 dilution.<br /><br />Lane 1: A549 cell lysate<br />Lane 2: A549 treated with 100nM Calyculin A for 30 minutes cell lysate<br />Lane 3: SH-SY5Y cell lysate<br />Lane 4: SH-SY5Y treated with 200nM Calyculin A and 1μM Okadaic Acid for 1 hour cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 51 kDa<br />Observed band size: 51 kDa<br /><br />Exposure time: 1 minute; ECL: K1801;<br />4-20% 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 (<a href="/products/HA723069" style="font-weight: bold;text-decoration: underline;">HA723069</a>) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Cell treatment (CT)

    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.

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of Phospho-GSK3 alpha (S21) on different lysates with Rabbit anti-Phospho-GSK3 alpha (S21) antibody (<a href="/products/HA723069" style="font-weight: bold;text-decoration: underline;">HA723069</a>) at 1/2,000 dilution.<br /><br />Lane 1: NIH/3T3 starved overnight cell lysate<br />Lane 2: NIH/3T3 starved overnight then treated with 100ng/mL PDGF for 5 minutes cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 51 kDa<br />Observed band size: 51 kDa<br /><br />Exposure time: 3 minutes; ECL: K1801;<br />4-20% 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 (<a href="/products/HA723069" style="font-weight: bold;text-decoration: underline;">HA723069</a>) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Cell treatment (CT)

    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.

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-Phospho-GSK3 alpha (S21) antibody (<a href="/products/HA723069" style="font-weight: bold;text-decoration: underline;">HA723069</a>) at 1/500 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA723069" style="font-weight: bold;text-decoration: underline;">HA723069</a>) 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.

    ☑ Cell treatment (CT)

    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.

  • Immunohistochemical analysis of paraffin-embedded human breast tissue with Rabbit anti-Phospho-GSK3 alpha (S21) antibody (<a href="/products/HA723069" style="font-weight: bold;text-decoration: underline;">HA723069</a>) at 1/200 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA723069" style="font-weight: bold;text-decoration: underline;">HA723069</a>) 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.

    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.

  • Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Rabbit anti-Phospho-GSK3 alpha (S21) antibody (<a href="/products/HA723069" style="font-weight: bold;text-decoration: underline;">HA723069</a>) at 1/200 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA723069" style="font-weight: bold;text-decoration: underline;">HA723069</a>) 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.

    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.

  • Dot blot analysis of Phospho-GSK3 alpha (S21) on different peptides with Rabbit anti-Phospho-GSK3 alpha (S21) antibody (<a href="/products/HA723069" style="font-weight: bold;text-decoration: underline;">HA723069</a>) at 1/1,000 dilution. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution for 1 hour at room temperature.<br /><br />Lane 1: Phospho-GSK3 alpha (S21) peptide (positive)<br />Lane 2: Unmodified GSK3 alpha peptide (negative)<br />Lane 3: Phospho-GSK3 beta (S9) peptide (negative)<br />Lane 4: Unmodified GSK3 beta peptide (negative)<br /><br />Proteins loading: 100ng, 25ng, 5ng;<br /><br />Blocking and dilution buffer: 5% NFDM/TBST;<br /><br />Exposure time: 10 seconds; ECL: K1801.

    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.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Citation

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