Immunohistochemistry analysis of Phospho-IRS1 [pY612] showing staining in the cytoplasm and nucleus of paraffin-embedded human skeletal muscle 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 [pY612] polyclonal antibody (44816G) diluted in 3% BSA-PBS at a dilution of 1:20 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|
|Published species reactivity||Rat, Human, Mouse, Not Applicable|
|Host / Isotype||Rabbit / IgG|
|Immunogen||The antiserum was produced against a chemically synthesized phosphopeptide derived from the region of human IRS-1 that contains tyrosine 612. The sequence is conserved in mouse, rat and chicken.|
|Purification||Antigen affinity chromatography|
|Storage buffer||Dulbecco's PBS, pH 7.3, with 1mg/ml BSA, 50% glycerol|
|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.
|Not Applicable||Not Cited||
Regulating the effects of GPR21, a novel target for type 2 diabetes.
44-816G was used in western blot to utilize a novel targe for type 2 diabetes and regulate the effects of GPR21
|Leonard S,Kinsella GK,Benetti E,Findlay JB||Scientific reports (6:null)||2016|
Severe insulin resistance alters metabolism in mesenchymal progenitor cells.
44-816G was used in immunocytochemistry and western blot to study Donohue syndrome in mesenchymal progenitor cells derived from human patients.
|Balhara B,Burkart A,Topcu V,Lee YK,Cowan C,Kahn CR,Patti ME||Endocrinology (156:2039)||2015|
Basal expression of insulin-like growth factor 1 receptor determines intrinsic resistance of cancer cells to a phosphatidylinositol 3-kinase inhibitor ZSTK474.
44-816G was used in western blot to examine the relationship of IGF1R and PI3Kis in chemotherapy resistance of cancer.
|Isoyama S,Kajiwara G,Tamaki N,Okamura M,Yoshimi H,Nakamura N,Kawamura K,Nishimura Y,Namatame N,Yamori T,Dan S||Cancer science (106:171)||2015|
Role of protein farnesylation in burn-induced metabolic derangements and insulin resistance in mouse skeletal muscle.
44-816G was used in western blot to elucidate the role of farnesylation in burn-induced metabolic aberration
|Nakazawa H,Yamada M,Tanaka T,Kramer J,Yu YM,Fischman AJ,Martyn JA,Tompkins RG,Kaneki M||PloS one (10:null)||2015|
The di-peptide Trp-His activates AMP-activated protein kinase and enhances glucose uptake independently of insulin in L6 myotubes.
44-816G was used in western blot to test if Trp-His activates AMP-activated protein kinase to regulate the glucose transport system in skeletal muscle.
|Soga M,Ohashi A,Taniguchi M,Matsui T,Tsuda T||FEBS open bio (4:898)||2014|
Salicylate acutely stimulates 5'-AMP-activated protein kinase and insulin-independent glucose transport in rat skeletal muscles.
44-816G was used in western blot to investigate the relationship between 5'-AMP-activated protein kinase and salicylate in skeletal muscle.
|Serizawa Y,Oshima R,Yoshida M,Sakon I,Kitani K,Goto A,Tsuda S,Hayashi T||Biochemical and biophysical research communications (453:81)||2014|
A novel inhibitor of the insulin/IGF signaling pathway protects from age-onset, neurodegeneration-linked proteotoxicity.
44-816G was used in western blot to evaluate a novel insulin/IGF signaling pathway inhibitor for its protective effect on age-onset, neurodegeneration-linked proteotoxicity
|El-Ami T,Moll L,Carvalhal Marques F,Volovik Y,Reuveni H,Cohen E||Aging cell (13:165)||2014|
Resistance exercise, but not endurance exercise, induces IKKß phosphorylation in human skeletal muscle of training-accustomed individuals.
44-816G was used in western blot to study the IκB kinase complex, TSC, MAPK, upstream Akt activators, and gene expression of selected cytokines in skeletal muscles.
|Møller AB,Vendelbo MH,Rahbek SK,Clasen BF,Schjerling P,Vissing K,Jessen N||Pflugers Archiv : European journal of physiology (465:1785)||2013|
Variant NKX3.1 and Serum IGF-1: Investigation of Interaction in Prostate Cancer.
44-816G was used in western blot to report that variant NKX3. cannot induce IGFBP-3 expression and that the NKX3. genotype does not modify the association between serum IGF-I levels and prostate cancer risk.
|Muhlbradt E,Ma J,Severi G,Ortner E,Hayes V,Hoang HN,Stampfer M,Giles G,Pollak M,Gelmann EP||Genes and cancer (4:535)||2013|
Obesity in mice with adipocyte-specific deletion of clock component Arntl.
44-816G was used in western blot to characterize transgenic mice with an adipocyte-specific deletion of Arntl.
|Paschos GK,Ibrahim S,Song WL,Kunieda T,Grant G,Reyes TM,Bradfield CA,Vaughan CH,Eiden M,Masoodi M,Griffin JL,Wang F,Lawson JA,Fitzgerald GA||Nature medicine (18:1768)||2012|
|Mouse||Not Cited||C1q/TNF-related protein-12 (CTRP12), a novel adipokine that improves insulin sensitivity and glycemic control in mouse models of obesity and diabetes.||Wei Z,Peterson JM,Lei X,Cebotaru L,Wolfgang MJ,Baldeviano GC,Wong GW||The Journal of biological chemistry (287:10301)||2012|
High IGF-IR activity in triple-negative breast cancer cell lines and tumorgrafts correlates with sensitivity to anti-IGF-IR therapy.
44-816G was used in western blot to examine the IGF signature in breast cancer cells after treatment wit anti-IGF-I receptor or insulin receptor inhibitors.
|Litzenburger BC,Creighton CJ,Tsimelzon A,Chan BT,Hilsenbeck SG,Wang T,Carboni JM,Gottardis MM,Huang F,Chang JC,Lewis MT,Rimawi MF,Lee AV||Clinical cancer research : an official journal of the American Association for Cancer Research (17:2314)||2011|
|Human||1:1000||Compensatory insulin receptor (IR) activation on inhibition of insulin-like growth factor-1 receptor (IGF-1R): rationale for cotargeting IGF-1R and IR in cancer.||Buck E,Gokhale PC,Koujak S,Brown E,Eyzaguirre A,Tao N,Rosenfeld-Franklin M,Lerner L,Chiu MI,Wild R,Epstein D,Pachter JA,Miglarese MR||Molecular cancer therapeutics (9:2652)||2010|
|Not Applicable||Not Cited||
NKX3.1 activates expression of insulin-like growth factor binding protein-3 to mediate insulin-like growth factor-I signaling and cell proliferation.
44-816G was used in western blot to study the role and function of NKX3.1 in prostate cells
|Muhlbradt E,Asatiani E,Ortner E,Wang A,Gelmann EP||Cancer research (69:2615)||2009|
|Rat||Not Cited||Insulin-like growth factor-I activation of Akt survival cascade in neuronal cells requires the presence of its cognate receptor in caveolae.||Lu X,Kambe F,Cao X,Yamauchi M,Seo H||Experimental cell research (314:342)||2008|
Type 1 Insulin-Like Growth Factor Receptor/Insulin Receptor Substrate 1 Signaling Confers Pathogenic Activity on Breast Tumor Cells Lacking REST.
44-816G was used in western blot to investigate how loss of RE-1 Silencing Transcription Factor affects breast cancer.
|Meyer K,Albaugh B,Schoenike B,Roopra A||Molecular and cellular biology (35:2991)||2015|
Insulin and IGF1 signalling pathways in human astrocytes in vitro and in vivo; characterisation, subcellular localisation and modulation of the receptors.
44-816G 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|
Myo1c regulates glucose uptake in mouse skeletal muscle.
44-816G was used in western blot to investigate the expression profile and role of Myo1c in skeletal muscle
|Toyoda T,An D,Witczak CA,Koh HJ,Hirshman MF,Fujii N,Goodyear LJ||The Journal of biological chemistry (286:4133)||2011|
||Insulin-like growth factor-I activation of Akt survival cascade in neuronal cells requires the presence of its cognate receptor in caveolae.||Lu X,Kambe F,Cao X,Yamauchi M,Seo H||Experimental cell research (314:342)||2008|