Immunohistochemistry analysis of Phospho-IRS1 [pS616] showing staining in the nucleus of paraffin-embedded human breast carcinoma tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a Phospho-IRS1 [pS616] polyclonal antibody (44550G) diluted in 3% BSA-PBS at a dilution of 1:100 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.
|Tested species reactivity||Human, Mouse, Rat|
|Published species reactivity||Human, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||The antiserum was produced against a chemically synthesized phosphopeptide derived from a region of IRS-1 that contains serine 616. The sequence is conserved in many species including human, mouse and rat.|
|Purification||Antigen affinity chromatography|
|Storage buffer||Dulbecco's PBS, pH 7.3, with 50% glycerol, 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1:20|
|Immunohistochemistry (Paraffin) (IHC (P))||1:10-1:100|
|Western Blot (WB)||Assay Dependent|
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.
Insulin and IGF1 signalling pathways in human astrocytes in vitro and in vivo; characterisation, subcellular localisation and modulation of the receptors.
44-550G was used in western blot to study the insulin/IGF1 signaling pathway in human astrocytes.
|Garwood CJ,Ratcliffe LE,Morgan SV,Simpson JE,Owens H,Vazquez-Villaseñor I,Heath PR,Romero IA,Ince PG,Wharton SB||Molecular brain (8:null)||2015|
Abnormal serine phosphorylation of insulin receptor substrate 1 is associated with tau pathology in Alzheimer's disease and tauopathies.
44-550G was used in immunohistochemistry - paraffin section to determine the expression of abnormal serine phosphorylation of insulin receptor substrate 1 in healthy and diseased human brains
|Yarchoan M,Toledo JB,Lee EB,Arvanitakis Z,Kazi H,Han LY,Louneva N,Lee VM,Kim SF,Trojanowski JQ,Arnold SE||Acta neuropathologica (128:679)||2014|
High fat diet produces brain insulin resistance, synaptodendritic abnormalities and altered behavior in mice.
44-550G was used in immunohistochemistry - paraffin section to determine how insulin resistance contributes to cognitive health.
|Arnold SE,Lucki I,Brookshire BR,Carlson GC,Browne CA,Kazi H,Bang S,Choi BR,Chen Y,McMullen MF,Kim SF||Neurobiology of disease (67:79)||2014|
|Human||Not Cited||Inhibition of FAK signaling activated by urokinase receptor induces dormancy in human carcinoma cells in vivo.||Aguirre Ghiso JA||Oncogene (21:2513)||2002|