Flow cytometry analysis of IRS1 was done on MCF7 cells. Cells were fixed with 70% ethanol for 10 minutes, permeabilized with 0.25% Triton™ X-100 for 20 minutes, and blocked with 5% BSA for 30 minutes at room temperature. Cells were labeled with IRS1 Rabbit Polyclonal Antibody (PA11058, red histogram) or with rabbit isotype control (pink histogram) at 3-5 ug/million cells in 2.5% BSA. After incubation at room temperature for 2 hours, the cells were labeled with Alexa Fluor® 488 Goat Anti-Rabbit Secondary Antibody (A11008) at a dilution of 1:400 for 30 minutes at room temperature. The representative 10,000 cells were acquired and analyzed for each sample using an Attune® Acoustic Focusing Cytometer. The purple histogram represents unstained control cells and the green histogram represents no-primary-antibody control.
|Tested species reactivity||Human, Mouse|
|Published species reactivity||Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues L(602) H T D D G Y M P M S P G V(615) of human IRS-1.|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Western Blot (WB)||2 ug/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Western Blot (WB)||See 1 publications below|
PA1-1058 detects a IRS-1 in mouse and human cells.
PA1-1058 has been successfully used in Western blot procedures. By Western blot this antibody detects an ~170 kDa protein representing IRS-1 in 3T3 L1 cell extract.
The PA1-1058 immunogen is a synthetic peptide corresponding to residues L(602) H T D D G Y M P M S P G V(615) of human IRS-1. The immunizing peptide is 92% conserved in rats, and mice. This peptide (Cat. # PEP-192) is available for use in neutralization and control experiments.
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 IRS-1 has been demonstrated to be a negative regulator of insulin signaling and is responsible for its degradation, although IRS-1 degradation pathways are not well understood. IRS-1 has also been shown to be constitutively activated in cancers such as breast cancer, Wilm and quote;s tumors, and adrenal cortical carcinomas, thus making IRS-1 phosphorylation and subsequent degradation an attractive therapeutic option.To date there have been four subtypes identified: IRS-1,2,3, and 4, with IRS-1 being widely expressed.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
The hyperglycemia-induced inflammatory response in adipocytes: the role of reactive oxygen species.
PA1-1058 was used in western blot to study the important role for ROS production in adipocytes and the associated insulin resistance and inflammatory response
|Lin Y,Berg AH,Iyengar P,Lam TK,Giacca A,Combs TP,Rajala MW,Du X,Rollman B,Li W,Hawkins M,Barzilai N,Rhodes CJ,Fantus IG,Brownlee M,Scherer PE||The Journal of biological chemistry (280:4617)||2005|