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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|
|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.
|Western Blot (WB)||See 12 publications below|
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"e;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.
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 IKKβ 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||Pflu¿gers 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 & 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|
Insulin receptor substrate-1; IRS-1; IRS1