44-550G

antibody from Invitrogen Antibodies

Targeting: IRS1 HIRS-1

Western blot (Supportive data in Antibodypedia)

Immunohistochemistry (Data presented on provider website)

Flow cytometry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

44-550G

Provider

Invitrogen Antibodies

Product name

Phospho-IRS1 (Ser616) Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

Purified from rabbit serum by sequential epitope-specific chromatography, this product contains enough material for 10 mini-blots. This antibody has been negatively preadsorbed using a non-phosphopeptide corresponding to the site of phosphorylation to remove antibody that is reactive with non-phosphorylated insulin receptor substrate-1 (IRS-1). The final product is generated by affinity chromatography using IRS-1 derived peptide that is phosphorylated at serine 616. The antibody has been used in western blotting. Previous lots of this antibody have been used in immunohistochemistry. In western analysis the positive control used was human embryonic kidney cell line transfected with a vector encoding IRS-1 (293T) and stimulated with a phorbol ester, TPA.

Reactivity

Human, Mouse, Rat

Host

Rabbit

Isotype

IgG

Vial size

100 µL

Storage

-20°C

References

Submitted references

Induction of Brain Insulin Resistance and Alzheimer's Molecular Changes by Western Diet.

Mietelska-Porowska A, Domańska J, Want A, Więckowska-Gacek A, Chutorański D, Koperski M, Wojda U
International journal of molecular sciences 2022 Apr 25;23(9)

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Altered Insulin Receptor Substrate 1 Phosphorylation in Blood Neuron-Derived Extracellular Vesicles From Patients With Parkinson's Disease.

Chou SY, Chan L, Chung CC, Chiu JY, Hsieh YC, Hong CT
Frontiers in cell and developmental biology 2020;8:564641

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Central insulin dysregulation and energy dyshomeostasis in two mouse models of Alzheimer's disease.

Velazquez R, Tran A, Ishimwe E, Denner L, Dave N, Oddo S, Dineley KT
Neurobiology of aging 2017 Oct;58:1-13

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Insulin and IGF1 signalling pathways in human astrocytes in vitro and in vivo; characterisation, subcellular localisation and modulation of the receptors.

Garwood CJ, Ratcliffe LE, Morgan SV, Simpson JE, Owens H, Vazquez-Villaseñor I, Heath PR, Romero IA, Ince PG, Wharton SB
Molecular brain 2015 Aug 22;8:51

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Abnormal serine phosphorylation of insulin receptor substrate 1 is associated with tau pathology in Alzheimer's disease and tauopathies.

Yarchoan M, Toledo JB, Lee EB, Arvanitakis Z, Kazi H, Han LY, Louneva N, Lee VM, Kim SF, Trojanowski JQ, Arnold SE
Acta neuropathologica 2014 Nov;128(5):679-89

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High fat diet produces brain insulin resistance, synaptodendritic abnormalities and altered behavior in mice.

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 2014 Jul;67:79-87

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Inhibition of FAK signaling activated by urokinase receptor induces dormancy in human carcinoma cells in vivo.

Aguirre Ghiso JA
Oncogene 2002 Apr 11;21(16):2513-24

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Extracts of 293T cells transfected with wild-type human IRS-1 and treated with 100 ng/mL TPA for 30 minutes were resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to PVDF.

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometry analysis of IRS1 [pS616] was done on MCF7 cells treated with Insulin (100nM, 10 minutes). Cells were fixed with 70% ethanol for 10 minutes, permeabilized with 0.25% Triton™ X-100 for 20 minutes, and blocked with 5% BSA for 30 minutes at room temperature. Cells were labeled with IRS1 [pS616] Rabbit Polyclonal Antibody (44550G, red histogram) or with rabbit isotype control (pink histogram) at 3-5 ug/million cells in 2.5% BSA. After incubation at room temperature for 2 hours, the cells were labeled with Alexa Fluor® 488 Goat Anti-Rabbit Secondary Antibody (A11008) at a dilution of 1:400 for 30 minutes at room temperature. The representative 10,000 cells were acquired and analyzed for each sample using an Attune® Acoustic Focusing Cytometer. The purple histogram represents unstained control cells and the green histogram represents no-primary-antibody control.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

NULL

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

NULL

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

NULL

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

NULL

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Fig. 4 Localisation of signalling components in human astrocytes. a Immunocytochemistry demonstrates nuclear localisation of IRS1 phosphorylated at serine 616 (S616) and S636/639 but not total IRS1 b Representative immunoblots of fractionated cell lysates showing the localisation of total IRS1 and IRS1 phosphorylated at S616 and tyrosine 612 (Y612) by immunoblot. Loading control for cytosplasmic fraction are EEF2 and for nuclear fraction is SP1. Molecular weight markers are indicated (kDa)