Phospho-IRS1 (S616) Antibody

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Description

R&D Systems MAB39783 (Mouse Monoclonal)

  • Host Species: Mouse

  • Clonality: Monoclonal (Clone #738710)

  • Target: Human IRS1 phosphorylated at S616

  • Applications: Western blotting, immunoblotting

  • Validation: Tested in MCF-7 cells treated with insulin and CIP, showing a specific band at ~150 kDa under reducing conditions .

  • Cross-Reactivity: Human, mouse, rat .

Thermo Fisher 44-550G (Rabbit Polyclonal)

  • Host Species: Rabbit

  • Target: Human IRS1 phosphorylated at S616

  • Applications: Western blotting, immunohistochemistry (previous lots)

  • Validation: Used in 293T cells transfected with IRS1 and treated with TPA .

  • Preadsorption: Negatively preadsorbed to remove reactivity with non-phosphorylated IRS1 .

Table 1: Antibody Comparison

FeatureR&D Systems MAB39783Thermo Fisher 44-550G
HostMouseRabbit
ClonalityMonoclonalPolyclonal
Species ReactivityHuman, mouse, ratHuman
ApplicationsWestern blotWestern blot, IHC
PreadsorptionNot specifiedYes

IRS1 Function and S616 Phosphorylation

IRS1 is a 150–180 kDa substrate of insulin/IGF-I receptors, mediating downstream signaling via PI-3 kinase and GRB2 . Phosphorylation at S616 is induced by insulin/IGF-I and angiotensin II, and is associated with insulin resistance .

Mechanism of S616 Phosphorylation

  • S6K1 Kinase: The mTOR/S6K1 pathway directly phosphorylates IRS1 at S616 in vitro, promoting its depletion from specific intracellular pools .

  • Pathological Role: S616 phosphorylation correlates with IRS1 degradation in tuberous sclerosis complex (TSC) models, contributing to insulin resistance .

Table 2: Key Research Findings

StudyKey Observations
R&D Systems Detects S616 phosphorylation in insulin-treated MCF-7 cells.
Thermo Fisher Validated in IRS1-transfected 293T cells treated with TPA.
Ouyang et al. (2006) S6K1 phosphorylates IRS1 at S616, linking mTOR signaling to insulin resistance.

Western Blotting

  • Sample Preparation: Use lysates from insulin/IGF-I-stimulated cells (e.g., MCF-7, 293T).

  • Optimization: Dilute antibody to 1 µg/mL (R&D Systems) or as recommended (Thermo Fisher).

  • Running Conditions: SDS-PAGE under reducing conditions for R&D Systems; exact conditions vary by protocol .

Clinical and Therapeutic Implications

S616 phosphorylation is a biomarker for insulin resistance in diseases like type 2 diabetes and TSC . Antibodies targeting this site enable quantitative assessment of IRS1 dysfunction, aiding in mechanistic studies and therapeutic development (e.g., mTOR inhibitors) .

This antibody is a critical tool for elucidating IRS1 signaling dynamics, with applications spanning basic research to translational medicine. Its specificity and validation across diverse models underscore its utility in advancing diabetes and metabolism research.

Product Specs

Buffer
Liquid in PBS containing 50% glycerol, 0.5% BSA, and 0.02% sodium azide.
Form
Liquid
Lead Time
Typically, we can ship products within 1-3 business days after receiving your order. Delivery times may vary depending on the purchase method or location. Please consult your local distributor for specific delivery times.
Synonyms
HIRS 1 antibody; HIRS1 antibody; Insulin receptor substrate 1 antibody; IRS 1 antibody; IRS-1 antibody; IRS1 antibody; IRS1_HUMAN antibody; OTTHUMP00000164234 antibody
Target Names
Uniprot No.

Target Background

Function
Phospho-IRS1 (S616) antibody may mediate the control of various cellular processes by insulin. Upon phosphorylation by the insulin receptor, it binds specifically to various cellular proteins containing SH2 domains such as the phosphatidylinositol 3-kinase p85 subunit or GRB2. When bound to the regulatory p85 subunit, it activates phosphatidylinositol 3-kinase.
Gene References Into Functions
  1. ER and IRS-1 subgroups appear to be critical factors for predicting breast cancer recurrence. In particular, patients with ER-positive and IRS-1-negative breast cancer may have a poorer prognosis and require more aggressive treatment. PMID: 29970713
  2. Research suggests that overexpression of LncRNA H19 could inhibit cell proliferation and promote apoptosis via downregulation of IRS-1 in thyroid cancer cells in vitro. These findings may provide valuable insights for understanding the LncRNA H19 functional network in thyroid cancer. Long noncoding RNA H19 could be a potential new target for antitumor therapy of thyroid cancer. PMID: 29332545
  3. Acute loss of IRS1/IRS2 or inhibition of IR/IGF1R in KRAS-mutant human NSCLC cells decreases amino acid uptake and lowers intracellular amino acid levels, while enhancing basal autophagy and sensitivity to autophagy and proteasome inhibitors. PMID: 29610318
  4. S6K1-dependent IRS-1pSer suppresses insulin signaling, leading to insulin resistance, a common occurrence in AD brains. Notably, miR-200b/c transfection of SH-SY5Y cells reduced the levels of IRS-1pSer. This finding indicates that miR-200b/c has the potential to alleviate insulin resistance by modulating S6K1. PMID: 29738527
  5. Collectively, miR-145 mimics suppress cell proliferation by targeting and inhibiting IRS1 expression, which in turn inhibits MAPK/ERK signaling pathways. PMID: 27799458
  6. Studies indicate that Y537S/D538G ESR1 mutant breast cancer cell lines exhibit enhanced proliferation in response to IGF1/IGF1R signaling, involving IRS1. Knockdown of IRS1 attenuates this enhanced IGF1/IGF1R signaling response in ESR1 mutant cells. (ESR1 = estrogen receptor 1; IGF1 = insulin like growth factor 1; IGF1R = IGF1 receptor; IRS1 = insulin receptor substrate 1) PMID: 29029116
  7. The effects of PF may be associated with its role in inhibiting de novo lipid synthesis and regulating the ROCK/IRS/Akt signaling pathways. PMID: 28380411
  8. Gene expression for insulin receptor substrate 1 (IRS-1), protein kinase B (Akt-2), and glucose transporter 4 (GLUT-4) genes were evaluated using real-time PCR. PMID: 28364599
  9. Reduced insulin receptor substrate-1 (IRS-1) staining in lung adenocarcinoma tissue microarray displayed a significant survival disadvantage, particularly within the Kirsten rat sarcoma viral oncogene homolog (KRAS) mutant subgroup. PMID: 27439864
  10. This study examined the normal regional brain expression of IRS-1 and found a significant correlation with the volumetric associated with regional atrophy in Alzheimer's disease. PMID: 28105773
  11. Data suggests that MIR126 induces autophagic flux in malignant mesothelioma (MM) cells by downregulating insulin receptor substrate-1 (IRS1) and disrupting the IRS1 signaling pathway. PMID: 27119351
  12. The IRS1/beta-Catenin Axis is Activated and Induces MYC Expression in Acute Lymphoblastic Leukemia Cells PMID: 27987331
  13. These effects were exerted by changes in the phosphorylation of IRS-1. PMID: 28011403
  14. Allele and genotype frequencies of rs1801278 in IRS1 showed significant differences between cases and controls for obstructive sleep apnea risk in the Chinese Han population. PMID: 27509181
  15. Gly972Arg of IRS-1 polymorphisms are associated with polycystic ovary syndrome. PMID: 27785750
  16. The G allele of the rs7578326 SNP in the LOC646736/IRS1 region is significantly associated with gestational diabetes mellitus. PMID: 28072873
  17. The results suggest that high glucose compromises the insulin signaling pathway in the glomerulus, promoting a proapoptotic environment. A possible critical step for this malfunction may lie at the level of IRS-1 phosphorylation. PMID: 27434075
  18. The C-allele of IRS1 variant rs2943650 was significantly associated with higher Body Fat Percentage overall and was significantly associated with lower levels of fasting insulin, homeostatic model assessment of insulin resistance, hemoglobin A1c, and triglycerides, and higher high-density lipoprotein cholesterol in United States Hispanics/Latinos. PMID: 27663718
  19. IRS-1 and IRS-2 signaling interaction with the microtubule cytoskeleton and its response to AKT determines the response to microtubule disruption in breast carcinoma cells. PMID: 28320862
  20. Consistent with these observations, LPIN1 levels were positively correlated with IRS1 expression in human breast cancer. Our results indicate a mechanism by which IRS1 expression is increased in breast cancer, and LPIN1 may be a promising drug target for anticancer therapy. PMID: 27729374
  21. IRS1 Gene Polymorphism is associated with Autism Spectrum Disorder. PMID: 27483248
  22. High IRS1 expression is associated with hepatocellular carcinoma. PMID: 27542674
  23. In the renal proximal tubule, insulin signaling via IRS1 is inhibited, while insulin signaling via IRS2 is preserved. Insulin signaling via IRS2 continues to stimulate sodium reabsorption in the proximal tubule and causes sodium retention, edema, and hypertension. PMID: 27247938
  24. miR-195 inhibits tumor angiogenesis by suppressing the IRS1-VEGF axis. PMID: 27133044
  25. The Arg(972) IRS-1 polymorphism is associated with increased risk and disease activity/severity of rheumatoid arthritis, and therefore may be a potential prognostic factor for RA. PMID: 25424426
  26. In this exploratory analysis, IRS1, ENNP1, and TRIB3, known to be associated with type 2 diabetes and harboring genes playing a prominent role in mediating insulin signaling, may modulate a number of cardiometabolic phenotypes in patients of Italian ancestry with newly-diagnosed type 2 diabetes. PMID: 26868433
  27. In accordance with previous studies, our findings suggest that the IRS1 G972R R allele and RR+GR genotype have protective effects for colorectal cancer (CRC) in overweight/obese patients and for obesity in patients with CRC. PMID: 26349669
  28. FRET-based translocation assays reveal that insulin promotes the association of both p62 and aPKC with the insulin-regulated scaffold IRS-1. PMID: 27143478
  29. HCV NS5A favors serine phosphorylation of IRS-1, promoting insulin resistance through IRS-1 serine phosphorylation and increased gluconeogenesis. PMID: 26604643
  30. High IRS1 Expression is associated with Colorectal Cancer. PMID: 26577117
  31. Failed to find any association between the IRS1 Gly972Arg polymorphism and T2DM. PMID: 26620983
  32. rs1801278 in IRS1 gene may play a role in type 2 diabetes risk, especially in Asian populations, and rs2943641 may be associated with type 2 diabetes risk in Caucasian populations. [meta-analysis] PMID: 26582067
  33. Upregulation of IRS1 was associated with metastasis of gastric carcinoma. PMID: 26684358
  34. The G allele of rs13431554 in the IRS-1 gene was associated with a hyperreactive platelet phenotype in coronary artery disease patients with T2DM. PMID: 27005817
  35. miR-126 functions as a tumor suppressor in glioma cells by targeting IRS-1 expression via the PI3K/AKT signaling pathways. PMID: 26617742
  36. Results showed that miR-128 was negatively associated with IRS1 in colorectal carcinoma (CRC) tissues and suggested that miR-128 serves as a tumor suppressor and blocks CRC growth and metastasis by targeting IRS1. PMID: 26352220
  37. IRS-specific gene signatures represent accurate surrogates of IGF activity and could predict response to anti-IGF therapy in breast cancer. PMID: 26991655
  38. Data suggest that IRS1 tyrosine phosphorylation, insulin sensitivity, and glucose internalization in visceral adipocytes can be up-regulated by dietary components (in this case, protocatechuic acid, a metabolite of dietary anthocyanins). PMID: 25944785
  39. Genetic association studies in a population in Austria: Data suggest that a haplotype upstream of IRS1 protects against insulin resistance, type 2 diabetes, dyslipidemias, and atherosclerosis. PMID: 26090471
  40. Arg972 IRS-1 inhibits endothelial nitric oxide synthase expression in human endothelial cells by upregulating miR-155 expression through the impairment of phosphatidylinositol-3 kinase signaling. PMID: 25902041
  41. Alpha-Syn overexpression negatively regulated IRS-1 via mTORC1/S6K1 signaling while activation of PP2A reverses this process. PMID: 25813876
  42. Hepatic insulin resistance in human obesity is: advanced; BMI-correlated; and involves aPKC-activating ceramide; aPKC levels and activity; IRS-1 levels, Akt activity, and FoxO1 phosphorylation; and increases in expression/abundance of PGC-1alpha. PMID: 26386696
  43. Arg972 IRS-1 enhances TNF-alpha-induced apoptosis in osteoblasts from rheumatoid arthritis patients. PMID: 25760103
  44. Hepatitis C virus infection suppresses the insulin signaling pathway and promotes insulin resistance by repressing PTEN, subsequently leading to decreased levels of IRS-1 and increased levels of Ser307-phosphorylated IRS-1. PMID: 25645159
  45. AFB1 downregulates IRS1 but paradoxically upregulates IRS2 through positive regulation of the stability of IRS2 and the proteasomal degradation of IRS1 in lung cancer cell lines A549 and SPCA-1. PMID: 25820822
  46. Gly972Arg was not associated with obesity, insulin resistance/sensitivity, or type 2 diabetes mellitus. PMID: 25214251
  47. Results show that in esophageal squamous cell carcinoma (ESCC), miR-126 was downregulated, and IRS-1 and GOLPH3, overexpressed, suggesting a tumor suppression role of miR-126 via the regulation of IRS-1 and GOLPH3. PMID: 25017784
  48. Our study found that the genetic polymorphisms rs10830963 and rs1387153 in MTNR1B and rs1801278 in IRS1 were associated with an increased risk of developing GDM. PMID: 25146448
  49. Results suggest that a genetic variation in the insulin signaling pathway genes IRS1, IRS2, and INSR may affect the therapeutic response of temporal lobe epilepsy. PMID: 25458098
  50. N-myristoylated Cblin prevents DEX-induced skeletal muscle atrophy in vitro and in vivo, and N-myristoylated Cblin more effectively prevents muscle atrophy than unmodified Cblin. PMID: 25689493

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Database Links

HGNC: 6125

OMIM: 125853

KEGG: hsa:3667

STRING: 9606.ENSP00000304895

UniGene: Hs.471508

Involvement In Disease
Diabetes mellitus, non-insulin-dependent (NIDDM)

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Applications : Immunohistochemical staining

Sample type: tissue

Review: Effects of n3-PUFAs (n-3, 300 mg/kg) and metformin (Met, 150 mg/kg) on relative hepatic tissue expression of insulin receptor substrate 1 (IRS-1) and phosphorylated insulin receptor substrate 1 (p-IRS1Ser 616) in high-fat diet/low dose streptozotocin (HFD-STZ)-induced NAFLD in rats by immunohistochemistry. (×400, scale bar = 50um).

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