IRS1 Recombinant Rabbit Monoclonal Antibody [SD2046]
Catalog# ET1612-45
IRS1 Recombinant Rabbit Monoclonal Antibody [SD2046]
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WB
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IF-Cell
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IF-Tissue
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IHC-P
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Human
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unconjugated
Overview
Product Name
IRS1 Recombinant Rabbit Monoclonal Antibody [SD2046]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic peptide within human IRS1 aa 1160-1242.
Species Reactivity
Human
Validated Applications
WB, IF-Cell, IF-Tissue, IHC-P
Target Molecular Weight
132 kDa
Positive Control
SW480, Hela, MCF-7, human breast carcinoma tissue.
Conjugation
unconjugated
Clone Number
SD2046
RRID
Reactivity Data
Verified (internally validated)
Reported in literature (not internally validated)
Predicted reactive (based on sequence homology)
Not recommended (failed internal validation)
| WB | IHC-P | FC | IF-Cell | |
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| human |
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| mouse |
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| rat |
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Product Features
Form
Liquid
Concentration
1 mg/mL.(The concentration of this product may be batch-dependent)
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). 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
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WB
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1:1,000
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IF-Cell
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1:50-1:200
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IF-Tissue
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1:50-1:200
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IHC-P
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1:50-1:200
Target
Function
The insulin receptor substrate-1 (IRS-1), a protein major substrate of the insulin receptor, is phosphorylated in response to stimulation of cells by insulin, insulin-like growth factor 1 (IGF-1) and interleukin 4 (IL-4). IRS-1 is phosphorylated on serine, threonine and tyrosine residues in a variety of tissues. An insulin-sensitive serine/threonine kinase casein kinase II mediates a portion of the insulin-stimulated serine/threonine phosphorylation of overexpressed IRS-1 in vivo. Thr 502 is identified as the major casein kinase II-catalyzed phosphorylation site in rat IRS-1, and Ser 99 is an additional phosphorylation site catalyzed by casein kinase II. Thus, casein kinase II-catalyzed phosphorylation of IRS-1 may be a component of the intracellular insulin signaling cascade. IRS-1 contains three putative binding sites for 14-3-3 (Ser 270, Ser 374 and Ser 641) and the motif around Ser 270 is located in the phosphotyrosine binding domain of IRS-1, which is responsible for the interaction with the insulin receptor. The association of 14-3-3 with IRS-1 increases significantly upon treatment with okadaic acid, a potent serine/ threonine phosphatase inhibitor. Therefore, the association of 14-3-3 protein may play a role in the regulation of insulin sensitivity by interrupting the association between the insulin receptor and IRS-1.
Background References
1. Lovly CM et al. Rationale for co-targeting IGF-1R and ALK in ALK fusion-positive lung cancer. Nat Med 20:1027-34 (2014).
2. Pan T et al. Cytohesins/ARNO: the function in colorectal cancer cells. PLoS One 9:e90997 (2014).
Post-translational Modification
Serine phosphorylation of IRS1 is a mechanism for insulin resistance. Ser-312 phosphorylation inhibits insulin action through disruption of IRS1 interaction with the insulin receptor (By similarity). Phosphorylation of Tyr-896 is required for GRB2-binding (By similarity). Phosphorylated by ALK. Phosphorylated at Ser-270, Ser-307, Ser-636 and Ser-1101 by RPS6KB1; phosphorylation induces accelerated degradation of IRS1. Phosphorylated on tyrosine residues in response to insulin. In skeletal muscles, dephosphorylated on Tyr-612 by TNS2 under anabolic conditions; dephosphorylation results in the proteasomal degradation of IRS1.; Ubiquitinated by the Cul7-RING(FBXW8) complex in a mTOR-dependent manner, leading to its degradation: the Cul7-RING(FBXW8) complex recognizes and binds IRS1 previously phosphorylated by S6 kinase (RPS6KB1 or RPS6KB2).
Subcellular Location
Cytoplasm, Nucleus.
UNIPROT
Synonyms
HIRS 1 antibody
HIRS1 antibody
Insulin receptor substrate 1 antibody
IRS 1 antibody
IRS-1 antibody
IRS1 antibody
IRS1_HUMAN antibody
OTTHUMP00000164234 antibody
Images
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ICC staining of IRS1 in SW480 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 (ET1612-45, 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).
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ICC staining of IRS1 in Hela 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 (ET1612-45, 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).
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ICC staining of IRS1 in MCF-7 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 (ET1612-45, 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).
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Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using anti-IRS1 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 (ET1612-45, 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.
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Flow cytometric analysis of HeLa cells labeling IRS1.
Cells were fixed and permeabilized. Then stained with the primary antibody (ET1612-45, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
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