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Glycogen synthase 1 / GYS1 Recombinant Rabbit Monoclonal Antibody [SN75-05]

Usd: 385

Specification

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Catalog# ET1611-59

Glycogen synthase 1 / GYS1 Recombinant Rabbit Monoclonal Antibody [SN75-05]

Application

  • WB

  • IF-Tissue

  • IHC-P

  • IP

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

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

Overview

Product Name

Glycogen synthase 1 / GYS1 Recombinant Rabbit Monoclonal Antibody [SN75-05]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within Human GYS1 aa 638-737 / 737.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Tissue, IHC-P, IP

Molecular Weight

Predicted band size: 84 kDa

Positive Control

HEK-293 cell lysate, A431 cell lysate, RD cell lysate, Mouse heart tissue lysate, Mouse skeletal muscle tissue lysate, Rat heart tissue lysate, Rat skeletal muscle tissue lysate, human skeletal muscle tissue, mouse cardiac muscle tissue, rat cardiac muscle tissue.

Conjugation

unconjugated

Clone Number

SN75-05

RRID

AB_3070038

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

  • 1:2,000

  • IF-Tissue

  • 1:50-1:200

  • IHC-P

  • 1:1,000

  • IP

  • 1-2μg/sample

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 α-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. Rubio-Villena C et al. Glycogenic activity of R6, a protein phosphatase 1 regulatory subunit, is modulated by the laforin-malin complex. Int J Biochem Cell Biol 45:1479-88 (2013).

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

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

Cytosol, Inclusion body, Membrane.

UNIPROT

SwissProt: P13807 Human

SwissProt: Q9Z1E4 Mouse

SwissProt: A2RRU1 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

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of Glycogen synthase 1 / GYS1 on different lysates with Rabbit anti-Glycogen synthase 1 / GYS1 antibody (<a href="/products/ET1611-59" style="font-weight: bold;text-decoration: underline;">ET1611-59</a>) at 1/1,000 dilution.<br /><br />Lane 1: HAP1-parental cell lysate<br />Lane 2: HAP1-Glycogen synthase 1 / GYS1 KD cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 81 kDa<br />Observed band size: 81 kDa<br /><br />Exposure time: 60 seconds; ECL: K1802;<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/ET1611-59" style="font-weight: bold;text-decoration: underline;">ET1611-59</a>) at 1/1,000 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.

    ☑ Knockdown (KD)

    Western blot analysis of Glycogen synthase 1 / GYS1 on different lysates with Rabbit anti-Glycogen synthase 1 / GYS1 antibody (ET1611-59) at 1/1,000 dilution.

    Lane 1: HAP1-parental cell lysate
    Lane 2: HAP1-Glycogen synthase 1 / GYS1 KD cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 81 kDa
    Observed band size: 81 kDa

    Exposure time: 60 seconds; ECL: K1802;
    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 (ET1611-59) at 1/1,000 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.

  • Western blot analysis of Glycogen synthase 1 / GYS1 on different lysates with Rabbit anti-Glycogen synthase 1 / GYS1 antibody (<a href="/products/ET1611-59" style="font-weight: bold;text-decoration: underline;">ET1611-59</a>) at 1/2,000 dilution.<br /><br />Lane 1: HEK-293 cell lysate<br />Lane 2: A431 cell lysate<br />Lane 3: RD cell lysate<br />Lane 4: Mouse heart tissue lysate<br />Lane 5: Mouse skeletal muscle tissue lysate<br />Lane 6: Rat heart tissue lysate<br />Lane 7: Rat skeletal muscle tissue lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 84 kDa<br />Observed band size: 84 kDa<br /><br />Exposure time: Lane 1-3: 46 seconds; Lane 4-7: 8 seconds; 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/ET1611-59" style="font-weight: bold;text-decoration: underline;">ET1611-59</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.

    Western blot analysis of Glycogen synthase 1 / GYS1 on different lysates with Rabbit anti-Glycogen synthase 1 / GYS1 antibody (ET1611-59) at 1/2,000 dilution.

    Lane 1: HEK-293 cell lysate
    Lane 2: A431 cell lysate
    Lane 3: RD cell lysate
    Lane 4: Mouse heart tissue lysate
    Lane 5: Mouse skeletal muscle tissue lysate
    Lane 6: Rat heart tissue lysate
    Lane 7: Rat skeletal muscle tissue lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 84 kDa
    Observed band size: 84 kDa

    Exposure time: Lane 1-3: 46 seconds; Lane 4-7: 8 seconds; 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 (ET1611-59) 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.

  • Immunohistochemical analysis of paraffin-embedded human skeletal muscle tissue with Rabbit anti-Glycogen synthase 1 / GYS1 antibody (<a href="/products/ET1611-59" style="font-weight: bold;text-decoration: underline;">ET1611-59</a>) at 1/1,000 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/ET1611-59" style="font-weight: bold;text-decoration: underline;">ET1611-59</a>) at 1/1,000 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 skeletal muscle tissue with Rabbit anti-Glycogen synthase 1 / GYS1 antibody (ET1611-59) at 1/1,000 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 (ET1611-59) at 1/1,000 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 cardiac muscle tissue with Rabbit anti-Glycogen synthase 1 / GYS1 antibody (<a href="/products/ET1611-59" style="font-weight: bold;text-decoration: underline;">ET1611-59</a>) at 1/1,000 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/ET1611-59" style="font-weight: bold;text-decoration: underline;">ET1611-59</a>) at 1/1,000 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 cardiac muscle tissue with Rabbit anti-Glycogen synthase 1 / GYS1 antibody (ET1611-59) at 1/1,000 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 (ET1611-59) at 1/1,000 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 cardiac muscle tissue with Rabbit anti-Glycogen synthase 1 / GYS1 antibody (<a href="/products/ET1611-59" style="font-weight: bold;text-decoration: underline;">ET1611-59</a>) at 1/1,000 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/ET1611-59" style="font-weight: bold;text-decoration: underline;">ET1611-59</a>) at 1/1,000 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 cardiac muscle tissue with Rabbit anti-Glycogen synthase 1 / GYS1 antibody (ET1611-59) at 1/1,000 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 (ET1611-59) at 1/1,000 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.

  • Glycogen synthase 1 / GYS1 was immunoprecipitated from 0.2 mg A431 cell lysate with <a href="/products/ET1611-59" style="font-weight: bold;text-decoration: underline;">ET1611-59</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/ET1611-59" style="font-weight: bold;text-decoration: underline;">ET1611-59</a> at 1/2,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.<br /><br />Lane 1: A431 cell lysate (input)<br />Lane 2: <a href="/products/ET1611-59" style="font-weight: bold;text-decoration: underline;">ET1611-59</a> IP in A431 cell lysate<br />Lane 3: Rabbit IgG instead of <a href="/products/ET1611-59" style="font-weight: bold;text-decoration: underline;">ET1611-59</a> in A431 cell lysate<br /><br />Blocking/Dilution buffer: 5% NFDM/TBST<br />Exposure time: 10 seconds; ECL: K1801

    Glycogen synthase 1 / GYS1 was immunoprecipitated from 0.2 mg A431 cell lysate with ET1611-59 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using ET1611-59 at 1/2,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.

    Lane 1: A431 cell lysate (input)
    Lane 2: ET1611-59 IP in A431 cell lysate
    Lane 3: Rabbit IgG instead of ET1611-59 in A431 cell lysate

    Blocking/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