Anti-phospho-IRS1 (Tyr1179) antibody (bs-8709R)

Anti-phospho-IRS1 (Tyr1179) antibody

DATASHEET

Host:Rabbit

Target Protein:phospho-IRS1 (Tyr1179)

IR:Immunogen Range:LN(p-Y)ID

Clonality:Polyclonal

Isotype:IgG

Entrez Gene:3667

Swiss Prot:P35568

Source:KLH conjugated synthesised phosphopeptide derived from human IRS1 around the phosphorylation site of Ser1179:LN(p-Y)ID 

Purification:affinity purified by Protein A

Storage:Preservative: 15mM Sodium Azide, Constituents: 1% BSA, 0.01M PBS, pH 7.4 Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles.

Background:Insulin receptor substrates (IRS) are responsible for several insulin related activities, such as glucose homeostasis, cell growth, cell transformation, apoptosis and insulin signal transduction. Serine/threonine phosphorylation of IRS1 has been demonstrated to be a negative regulator of insulin signaling and is responsible for its degradation, although IRS1 degradation pathways are not well understood. IRS1 has also been shown to be constitutively activated in cancers such as breast cancer, Wilm's tumors, and adrenal cortical carcinomas, thus making IRS1 phosphorylation and subsequent degradation an attractive therapeutic target. To date there have been four subtypes identified: IRS1, 2, 3 and 4, with IRS1 being widely expressed. Function : May mediate the control of various cellular processes by insulin. When phosphorylated by the insulin receptor binds specifically to various cellular proteins containing SH2 domains such as phosphatidylinositol 3-kinase p85 subunit or GRB2. Activates phosphatidylinositol 3-kinase when bound to the regulatory p85 subunit.

Size:100ul

Concentration:1mg/ml

Applications:WB(1:500-2000)
ELISA(1:5000-10000)
IHC-P(1:100-500)
IHC-F(1:100-500)
Flow-Cyt(2ug/Test)
ICC(1:100-500)
IF(1:100-500)

Cross Reactive Species:Human
Mouse
Rat
Chicken
Dog
Pig
Cow
Rabbit
Sheep
Guinea Pig
Danio rerio,.

For research use only. Not intended for diagnostic or therapeutic use.

VALIDATION IMAGES

Sample:
A549 Cell (Human) Lysate at 40 ug
Hela Cell (Human) Lysate at 40 ug
MCF-7 Cell (Human) Lysate at 40 ug
Primary: Anti- phospho-IRS1 (Tyr1179) (bs-8709R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 132 kD
Observed band size: 132 kD

Paraformaldehyde-fixed, paraffin embedded (Mouse 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 (phospho-IRS1 (Tyr1179)) Polyclonal Antibody, Unconjugated (bs-8709R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Blank control: HepG2.
Primary Antibody (green line): Rabbit Anti-phospho-IRS1 (Tyr1179) antibody (bs-8709R)
Dilution: 2μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody : Goat anti-rabbit IgG-AF647
Dilution: 1μg /test.
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at -20℃. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.