AS160 Recombinant Rabbit Monoclonal Antibody [PSH10-95] - BSA and Azide free
Catalog# HA751385
AS160 Recombinant Rabbit Monoclonal Antibody [PSH10-95] - BSA and Azide free
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WB
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IHC-P
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IP
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Human
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Rat
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HA723262
含抗保成分
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unconjugated
Overview
Product Name
AS160 Recombinant Rabbit Monoclonal Antibody [PSH10-95] - BSA and Azide free
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
Product Features
Form
Liquid
Concentration
1mg/ml
Storage Instructions
Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage Buffer
1*PBS (pH7.4).
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:2,000-1:10,000
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IHC-P
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1:200-1:1,000
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IP
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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.
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
ExpandImages
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Western blot analysis of AS160 on different lysates with Rabbit anti-AS160 antibody (HA751385) 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 (HA751385) 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 (HA751385) 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 (HA751385) 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 (HA751385) 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 (HA751385) 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 (HA751385) 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 (HA751385) 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 (HA751385) 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 (HA751385) 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 (HA751385) 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 (HA751385) 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 HA751385 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA751385 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: HA751385 IP in HeLa cell lysate
Lane 3: Rabbit IgG instead of HA751385 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"