This gene encodes the insulin receptor substrate 2, a cytoplasmic signaling molecule that mediates effects of insulin, insulin-like growth factor 1, and other cytokines by acting as a molecular adaptor between diverse receptor tyrosine kinases and downstream effectors. The product of this gene is phosphorylated by the insulin receptor tyrosine kinase upon receptor stimulation, as well as by an interleukin 4 receptor-associated kinase in response to IL4 treatment. [provided by RefSeq, Jul 2008]
Function:
May mediate the control of various cellular processes by insulin.
Subunit:
Interacts with PHIP.
Subcellular Location:
Cytoplasm, cytosol.
Post-translational modifications:
Phosphorylated upon DNA damage, probably by ATM or ATR.
Similarity:
Contains 1 IRS-type PTB domain.
Contains 1 PH domain.
SWISS:
Q9Y4H2
Gene ID:
8660
Database links:
Entrez Gene: 8660 Human
Entrez Gene: 384783 Mouse
Entrez Gene: 29376 Rat
Omim: 600797 Human
SwissProt: Q9Y4H2 Human
SwissProt: P81122 Mouse
Unigene: 442344 Human
Unigene: 407207 Mouse
IRS-2是肝胰岛素信号转导核心介子，IRS-2基因的缺失或IRS-2信号网络上一些关键信号分子的异常改变，都将导致肝胰岛素信号转导能力减弱，出现肝胰岛素抵抗.
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Paraformaldehyde-fixed, paraffin embedded (Rat skeletal muscle); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (IRS-2) Polyclonal Antibody, Unconjugated (SL20748R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (Rat brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (IRS-2) Polyclonal Antibody, Unconjugated (SL20748R) at 1:500 overnight at 4°C, followed by a conjugated secondary (sp-0023) for 20 minutes and DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat kidney tissue); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (IRS-2) Polyclonal Antibody, Unconjugated (SL20748R) at 1:400 overnight at 4°C, followed by a conjugated secondary (sp-0023) for 20 minutes and DAB staining.
SH-SY5Y cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, SLC0005) at 37°C for 20 min; Antibody incubation with (IRS-2) polyclonal Antibody, Unconjugated (SL20748R) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.
Blank control (Black line): Raji (Black).
Primary Antibody (red line): Rabbit Anti-IRS-2 antibody (SL20748R)
Dilution: 1μg /10^6 cells;
Isotype Control Antibody (green line): Rabbit IgG .
Secondary Antibody (white blue line): Goat anti-rabbit IgG-PE
Dilution: 1μg /test.
Protocol
The cells were fixed with 4% PFA （10min）and then permeabilized with 0.1% PBS-Tween for 20 min at room temperature . Cells stained with Primary Antibody for 30 min at room temperature. The cells were then incubated in 1 X PBS/2%BSA/10% goat serum to block non-specific protein-protein interactions followed by the antibody for 15 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.
Blank control (black line): HepG2(black) (The cells were fixed with 2% paraformaldehyde (10 min) , then permeabilized with PBST for 30 min on room temperature)
Primary Antibody (Red line): Rabbit Anti-IRS-2 antibody (SL20748R) ;
Dilution: 1μg /10^6 cells;
Isotype Control Antibody (green line): Rabbit IgG .
Secondary Antibody (white blue line): Goat anti-rabbit IgG-FITC;Dilution: 1μg /test.
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