- 44-550G - Invitrogen Antibodies IRS1 antibody

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)

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

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

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

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

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

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

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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.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: 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 (Product # 44-550G) diluted in 3% BSA-PBS at a dilution of 1:100 overnight at 4ºC 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.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of Phospho-IRS1 (pS616) showing staining in the nucleus of paraffin-embedded mouse 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 (pS616) polyclonal antibody (Product # 44-550G) diluted in 3% BSA-PBS at a dilution of 1:20 overnight at 4ºC 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.

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.

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)

44-550G

Antibody data