Skip to content

AS160 Recombinant Rabbit Monoclonal Antibody [PSH10-95]

Usd: 385

Specification

Bulk Order Quote | Customization Request

Catalog# HA723262

AS160 Recombinant Rabbit Monoclonal Antibody [PSH10-95]

Application

  • WB

  • IHC-P

  • IP

Reactivity

  • Human

  • Rat

BSA and Azide free
Conjugation

  • unconjugated

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

Overview

Product Name

AS160 Recombinant Rabbit Monoclonal Antibody [PSH10-95]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within human AS160 aa 1-500.

Species Reactivity

Human, Rat

Validated Applications

WB, IHC-P, IP

Molecular Weight

Predicted band size: 147 kDa

Positive Control

HeLa cell lysate, HEK-293 cell lysate, HepG2 cell lysate, MCF7 cell lysate, Caco-2 cell lysate, human heart tissue, human skeletal muscle tissue, rat brain tissue, rat heart tissue, rat skeletal muscle tissue.

Conjugation

unconjugated

Clone Number

PSH10-95

Reactivity Data

WB IHC-P IP
Human
Tested Tested
Tested Tested
Tested Tested
Rat
Tested
Tested Tested
Mouse
Not Recommended
Not Recommended

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

  • 1:2,000-1:10,000

  • IHC-P

  • 1:200-1:1,000

  • IP

  • 1-2μg/sample

Target

Function

AS160 (Akt substrate of 160 kDa), which was originally known as TBC1 domain family member 4 (TBC1D4), is a Rab GTPase-activating protein that in humans is encoded by the TBC1D4 gene. The 160 kD protein product was first discovered in a screen for novel substrates of the serine-threonine kinase Akt2, which phosphorylates AS160 at Thr-642 and Ser-588 after insulin stimulation. Insulin stimulation of fat and muscle cells results in translocation of the glucose transporter GLUT4 to the plasma membrane, and this translocation process is dependent on phosphorylation of AS160. The role of AS160 in GLUT4 translocation is mediated by its GTPase activating domain and interactions with Rab proteins in vesicle formation, increasing GLUT4 translocation when its GTPase activity is inhibited by Akt phosphorylation. Specifically, this inhibition activates RAB2A, RAB8A, RAB10 and RAB14. AS160 also contains a calmodulin-binding domain, and this domain mediates phosphorylation-independent glucose uptake in muscle cells.

Background References

1. Sharma M et al. AKT ISOFORMS-AS160-GLUT4: The defining axis of insulin resistance. Rev Endocr Metab Disord. 2021 Dec

2. Su S et al. AS160 is a lipid-responsive regulator of cardiac Ca(2+) homeostasis by controlling lysophosphatidylinositol metabolism and signaling. Nat Commun. 2024 Nov

Subcellular Location

Cytoplasm.

UNIPROT

SwissProt: O60343 Human

Entrez Gene: 306117 Rat

Synonyms

Acrg embryonic lethality (mouse) minimal region ortholog antibody

Acrg embryonic lethality minimal region ortholog antibody

Acrg embryonic lethality mouse minimal region ortholog antibody

Akt substrate of 160 kDa antibody

AS 160 antibody

AS160 antibody

BUB2 antibody

CDC16 antibody

KIAA0603 antibody

NIDDM5 antibody

Expand

Images

  • Western blot analysis of AS160 on different lysates with Rabbit anti-AS160 antibody (<a href="/products/HA723262" style="font-weight: bold;text-decoration: underline;">HA723262</a>) at 1/2,000 dilution.<br /><br />Lane 1: HeLa cell lysate (20 µg/Lane)<br />Lane 2: HEK-293 cell lysate (20 µg/Lane)<br />Lane 3: HepG2 cell lysate (20 µg/Lane)<br />Lane 4: MCF7 cell lysate (20 µg/Lane)<br />Lane 5: Caco-2 cell lysate (20 µg/Lane)<br /><br />Predicted band size: 147 kDa<br />Observed band size: 160 kDa<br /><br />Exposure time: 10 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/HA723262" style="font-weight: bold;text-decoration: underline;">HA723262</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 AS160 on different lysates with Rabbit anti-AS160 antibody (HA723262) at 1/2,000 dilution.

    Lane 1: HeLa cell lysate (20 µg/Lane)
    Lane 2: HEK-293 cell lysate (20 µg/Lane)
    Lane 3: HepG2 cell lysate (20 µg/Lane)
    Lane 4: MCF7 cell lysate (20 µg/Lane)
    Lane 5: Caco-2 cell lysate (20 µg/Lane)

    Predicted band size: 147 kDa
    Observed band size: 160 kDa

    Exposure time: 10 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 (HA723262) 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 heart tissue with Rabbit anti-AS160 antibody (<a href="/products/HA723262" style="font-weight: bold;text-decoration: underline;">HA723262</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/HA723262" style="font-weight: bold;text-decoration: underline;">HA723262</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 heart tissue with Rabbit anti-AS160 antibody (HA723262) 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 (HA723262) 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-AS160 antibody (<a href="/products/HA723262" style="font-weight: bold;text-decoration: underline;">HA723262</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/HA723262" style="font-weight: bold;text-decoration: underline;">HA723262</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.

    Immunohistochemical analysis of paraffin-embedded human skeletal muscle tissue with Rabbit anti-AS160 antibody (HA723262) 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 (HA723262) 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 rat brain tissue with Rabbit anti-AS160 antibody (<a href="/products/HA723262" style="font-weight: bold;text-decoration: underline;">HA723262</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/HA723262" style="font-weight: bold;text-decoration: underline;">HA723262</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 brain tissue with Rabbit anti-AS160 antibody (HA723262) 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 (HA723262) 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 heart tissue with Rabbit anti-AS160 antibody (<a href="/products/HA723262" style="font-weight: bold;text-decoration: underline;">HA723262</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/HA723262" style="font-weight: bold;text-decoration: underline;">HA723262</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 heart tissue with Rabbit anti-AS160 antibody (HA723262) 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 (HA723262) 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 skeletal muscle tissue with Rabbit anti-AS160 antibody (<a href="/products/HA723262" style="font-weight: bold;text-decoration: underline;">HA723262</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/HA723262" style="font-weight: bold;text-decoration: underline;">HA723262</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-AS160 antibody (HA723262) 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 (HA723262) 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.

  • AS160 was immunoprecipitated from 0.2 mg HeLa cell lysate with <a href="/products/HA723262" style="font-weight: bold;text-decoration: underline;">HA723262</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/HA723262" style="font-weight: bold;text-decoration: underline;">HA723262</a> at 1/1,000 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: HeLa cell lysate (input)<br />Lane 2: <a href="/products/HA723262" style="font-weight: bold;text-decoration: underline;">HA723262</a> IP in HeLa cell lysate<br />Lane 3: Rabbit IgG instead of <a href="/products/HA723262" style="font-weight: bold;text-decoration: underline;">HA723262</a> in HeLa cell lysate<br /><br />Blocking/Dilution buffer: 5% NFDM/TBST<br />Exposure time: 9 seconds; ECL: <a href="/products/K1801" style="font-weight: bold;text-decoration: underline;">K1801</a>

    AS160 was immunoprecipitated from 0.2 mg HeLa cell lysate with HA723262 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA723262 at 1/1,000 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: HeLa cell lysate (input)
    Lane 2: HA723262 IP in HeLa cell lysate
    Lane 3: Rabbit IgG instead of HA723262 in HeLa cell lysate

    Blocking/Dilution buffer: 5% NFDM/TBST
    Exposure time: 9 seconds; ECL: K1801

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

Citation