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)||3-5 µg/million cells|
|Immunohistochemistry (Paraffin) (IHC (P))||1:10-1:100|
|Western Blot (WB)||Assay Dependent|
* 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.
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.
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|