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Phospho-Glycogen synthase 1 (S641) Recombinant Rabbit Monoclonal Antibody [SR46-06]

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Specification

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Catalog# ET1602-13

Phospho-Glycogen synthase 1 (S641) Recombinant Rabbit Monoclonal Antibody [SR46-06]

Application

  • WB

  • IF-Cell

  • IHC-P

  • IP

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

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

Overview

Product Name

Phospho-Glycogen synthase 1 (S641) Recombinant Rabbit Monoclonal Antibody [SR46-06]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic phospho-peptide corresponding to residues surrounding Ser641 of Human Glycogen synthase 1.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IHC-P, IP

Molecular Weight

Predicted band size: 84 kDa

Positive Control

SK-Br-3 cell lysate, A431 cell lysate, HepG2 cell lysate, Mouse liver tissue lysate, A549, NIH/3T3, mouse liver tissue, mouse skeletal muscle tissue, mouse smooth muscle tissue, rat skeletal muscle tissue.

Conjugation

unconjugated

Clone Number

SR46-06

RRID

AB_3069625

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

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

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:500

  • IF-Cell

  • 1:50

  • IHC-P

  • 1:50-1:200

  • IP

  • Use at an assay dependent concentration.

Target

Function

Glycogen [starch] synthase belongs to the mammalian/fungal glycogen synthase family of proteins. Two forms of this protein exist, a liver form and a muscle form, both of which have the same function in the glycogen biosynthesis pathway. Glycogen synthase transfers the glycosyl residue from UDP-Glucose to the nonreducing end of ?-1,4-glucan. The liver glycogen synthase protein is truncated by 34 amino acids compared to the muscle form. However, these enzymes differ significantly in their amino- and carboxyl-terminal regions. Muscle glycogen synthase serves to fuel muscular activity only and is regulated by muscle contraction and by catecholamines. Liver glycogen synthase mediates blood glucose homeostasis in response to nutritional cues. Defects in the gene encoding liver glycogen synthase results in glycogen storage disease type 0 (GSD0), a rare form of fasting ketotic hypoglycemia.

Background References

1. Zhang JS et al. Differential activity of GSK-3 isoforms regulates NF-kB and TRAIL- or TNFa induced apoptosis in pancreatic cancer cells. Cell Death Dis 5:e1142 (2014).

2. Reichelt ME et al. Myocardial glycophagy - A specific glycogen handling response to metabolic stress is accentuated in the female heart. J Mol Cell Cardiol 65C:67-75 (2013).

Sequence Similarity

Belongs to the glycosyltransferase 3 family.

Post-translational Modification

Phosphorylation at Ser-8 by AMPK inactivates the enzyme activity. Primed phosphorylation at Ser-657 (site 5) by CSNK2A1 and CSNK2A2 is required for inhibitory phosphorylation at Ser-641 (site 3a), Ser-645 (site 3b), Ser-649 (site 3c) and Ser-653 (site 4) by GSK3A an GSK3B (By similarity). Phosphorylated at Ser-641 by DYRK2, leading to inactivation (By similarity). Phosphorylated at Ser-641 by PASK, leading to inactivation; phosphorylation by PASK is inhibited by glycogen. Dephosphorylation at Ser-641 and Ser-645 by PP1 activates the enzyme.

Subcellular Location

Cytoplasm

UNIPROT

SwissProt: P13807 Human

SwissProt: Q9Z1E4 Mouse

Entrez Gene: 690987 Rat

Synonyms

Glycogen [starch] synthase antibody

Glycogen synthase 1 (muscle) antibody

Glycogen synthase 1 antibody

GSY antibody

GYS antibody

Gys1 antibody

GYS1_HUMAN antibody

muscle antibody

Images

  • Western blot analysis of Phospho-Glycogen synthase 1 (S641) on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (<a href="/products/ET1602-13" style="font-weight: bold;text-decoration: underline;">ET1602-13</a>, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:200,000 dilution was used for 1 hour at room temperature.<br />Positive control: <br />Lane 1: SK-Br-3 cell lysate<br />Lane 2: A431 cell lysate

    Western blot analysis of Phospho-Glycogen synthase 1 (S641) on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1602-13, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.
    Positive control:
    Lane 1: SK-Br-3 cell lysate
    Lane 2: A431 cell lysate

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of Phospho-Glycogen synthase 1 (S641) on HepG2 cell lysates.<br /><br />Lane 1: HepG2 cells, whole cell lysate, 10ug/lane<br />Lane 2: HepG2 cells treated with 2.8ug/ul lambda-PP for 30 minutes, whole cell lysates, 10ug/lane<br /><br />All lanes :<br />Anti-Phospho-Glycogen synthase 1 (S641) antibody (<a href="/products/ET1602-13" style="font-weight: bold;text-decoration: underline;">ET1602-13</a>) at 1/500 dilution. Anti-GAPDH antibody (<a href="/products/ET1601-4" style="font-weight: bold;text-decoration: underline;">ET1601-4</a>) at 1/10,000 dilution. Goat Anti-Rabbit IgG H&L (HRP) (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/200,000 dilution.<br /><br />Predicted band size: 84 kDa<br />Observed band size: 84 kDa<br /><br />Blocking and diluting buffer: 5% BSA.<br /><br />Exposure time: 1 minute 34 seconds

    ☑ Cell treatment (CT)

    Western blot analysis of Phospho-Glycogen synthase 1 (S641) on HepG2 cell lysates.

    Lane 1: HepG2 cells, whole cell lysate, 10ug/lane
    Lane 2: HepG2 cells treated with 2.8ug/ul lambda-PP for 30 minutes, whole cell lysates, 10ug/lane

    All lanes :
    Anti-Phospho-Glycogen synthase 1 (S641) antibody (ET1602-13) 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: 84 kDa
    Observed band size: 84 kDa

    Blocking and diluting buffer: 5% BSA.

    Exposure time: 1 minute 34 seconds

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of Phospho-Glycogen synthase 1 (S641) on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (<a href="/products/ET1602-13" style="font-weight: bold;text-decoration: underline;">ET1602-13</a>, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:5,000 dilution was used for 1 hour at room temperature.<br />Positive control: <br />Lane 1: Mouse liver tissue lysate, untreated <br />Lane 2: Mouse liver tissue lysate, treated with AP

    ☑ Cell treatment (CT)

    Western blot analysis of Phospho-Glycogen synthase 1 (S641) on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1602-13, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
    Positive control:
    Lane 1: Mouse liver tissue lysate, untreated
    Lane 2: Mouse liver tissue lysate, treated with AP

  • ICC staining of Phospho-Glycogen synthase 1 (S641) in A549 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 (<a href="/products/ET1602-13" style="font-weight: bold;text-decoration: underline;">ET1602-13</a>, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor&reg;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-Glycogen synthase 1 (S641) in A549 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 (ET1602-13, 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-Glycogen synthase 1 (S641) in NIH/3T3 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 (<a href="/products/ET1602-13" style="font-weight: bold;text-decoration: underline;">ET1602-13</a>, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor&reg;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-Glycogen synthase 1 (S641) in NIH/3T3 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 (ET1602-13, 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 mouse liver tissue using anti-Phospho-Glycogen synthase 1 (S641) 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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1602-13" style="font-weight: bold;text-decoration: underline;">ET1602-13</a>, 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 mouse liver tissue using anti-Phospho-Glycogen synthase 1 (S641) 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 (ET1602-13, 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 mouse skeletal muscle tissue with Rabbit anti-Phospho-Glycogen synthase 1 (S641) antibody (<a href="/products/ET1602-13" style="font-weight: bold;text-decoration: underline;">ET1602-13</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/ET1602-13" style="font-weight: bold;text-decoration: underline;">ET1602-13</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 mouse skeletal muscle tissue with Rabbit anti-Phospho-Glycogen synthase 1 (S641) antibody (ET1602-13) 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 (ET1602-13) 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 mouse smooth muscle tissue using anti-Phospho-Glycogen synthase 1 (S641) 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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1602-13" style="font-weight: bold;text-decoration: underline;">ET1602-13</a>, 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 mouse smooth muscle tissue using anti-Phospho-Glycogen synthase 1 (S641) 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 (ET1602-13, 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 rat skeletal muscle tissue with Rabbit anti-Phospho-Glycogen synthase 1 (S641) antibody (<a href="/products/ET1602-13" style="font-weight: bold;text-decoration: underline;">ET1602-13</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/ET1602-13" style="font-weight: bold;text-decoration: underline;">ET1602-13</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 rat skeletal muscle tissue with Rabbit anti-Phospho-Glycogen synthase 1 (S641) antibody (ET1602-13) 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 (ET1602-13) 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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Citation