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P irs1 s1101

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The P-IRS1 [S1101] is a laboratory reagent used to detect the phosphorylation of insulin receptor substrate 1 (IRS1) at serine 1101. It is a highly specific antibody that can be used in various analytical techniques, such as Western blotting, to study insulin signaling and related cellular processes.

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Lab products found in correlation

Ps473 akt, by Cell Signaling Technology (3 mentions) Hif 1α, by BD (2 mentions) Sc 44225, by Santa Cruz Biotechnology (2 mentions) Sc 35316, by Santa Cruz Biotechnology (2 mentions) Hif 1α oh p564, by Cell Signaling Technology (2 mentions) Lipofectamine 3000, by Thermo Fisher Scientific (2 mentions) Hif 1α, by Abcam (2 mentions) Ubiquitin, by Cell Signaling Technology (2 mentions) Phosphatase inhibitor cocktail 3, by Merck Group (2 mentions) Total akt, by Cell Signaling Technology (2 mentions) Ripa buffer, by Merck Group (2 mentions) Phosphatase inhibitor cocktail 2, by Merck Group (2 mentions) Protease inhibitor cocktail, by Merck Group (2 mentions) Gradient sds polyacrylamide gel, by Bio-Rad (2 mentions) Odyssey infrared imaging system, by LI COR (2 mentions) Pim447, by Selleck Chemicals (1 mentions) Hif1a, by Cell Signaling Technology (1 mentions) Wave2, by Cell Signaling Technology (1 mentions) Trypsin edta, by Corning (1 mentions) Dimethyl sulfoxide (dmso), by Thermo Fisher Scientific (1 mentions)

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P-IRS1 (25 citations)

6 protocols using p irs1 s1101

1

Hypoxia signaling protein regulation

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siHIF-1α experiments were performed using a forward transfection protocol (Lipofectamine 3000 Invitrogen). HIF-1α and siRNA was purchased from Santa Cruz Biotechnology (sc-44225 and sc-35316, respectively). PIM1 siRNA sequences were previously reported [58 ]. The following western blot antibodies were used for western blot: actin (BD Biosciences); and HA, HIF-1α (cat #: 3724S), HIF-1α-OH [P564] (cat #: 3434S), p-IRS1 [S1101] (cat #: 2385S), PHD2 (cat #: 4835S), PIM1 (cat #: 4835S), and ubiquitin (cat #: 3933S) (Cell Signaling Technologies). The p-HIF-1α [T455] antibody was generated by Pacific Immunology for western blot and IHC in this manuscript. The following antibodies were also used for IHC: CC3 (cat #: ab4051), CD31 (cat #: ab124432), HIF-1α (cat #: ab16066) and PIM1 (cat #: ab245417)(Abcam).
Casillas A.L., Chauhan S.S., Toth R.K., Sainz A.G., Clements A.N., Jensen C.C., Langlais P.R., Miranti C.K., Cress A.E, & Warfel N.A. (2021). Direct phosphorylation and stabilization of HIF-1α by PIM1 kinase drives angiogenesis in solid tumors. Oncogene, 40(32), 5142-5152.
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2

Comprehensive Protein Expression Analysis

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Trypsin-EDTA (Cat# 25-053-CI), PBS (Cat# 21-031-CV), and laminin (Cat# 354232) were purchased from Corning. Recombinant EGF was purchased from Life Technologies (Cat# PHG0311). IGF was purchased from Sigma-Aldrich (Cat# I3769). PIM447 was purchased from Selleck Chemicals (Cat# S7985). AZD1208 was acquired from AdooQ Bioscience (Cat# A13203-750). DMSO was purchased from Thermo Fisher Scientific (Cat# 97064-724). Cycloheximide was purchased from VWR (Cat# 97064-724), and doxycycline was purchased from Sigma-Aldrich (Cat# D9891-5G). Recombinant myelin basic protein (MBP) was a gift from Dr. Greg Rogers, and recombinant ABI2 protein was purchased from Origene (Cat# TP300637). Radio-labeled ATP was purchased from Perkin Elmer (Cat# BLU502A). The antibody to ABI2 was purchased from Bethyl Laboratories (Cat# A302-499A-M). GFP (Cat# 2956S), HA (Cat# 3724S), HIF-1a (Cat# 14179S), p-IRS1 [S1101] (Cat# 2385S), PIM1 (Cat# 3247S), and WAVE2 (Cat# 3659S) antibodies were purchased from Cell Signaling Technology. The antibody for actin was purchased from BD Biosciences (Cat# 61656). PIM1 (Cat# ab75776) and Ki67 (Cat#ab833) antibodies used for immunohistochemistry were purchased from Abcam.
Jensen C.C., Clements A.N., Liou H., Ball L.E., Bethard J.R., Langlais P.R., Toth R.K., Chauhan S.S., Casillas A.L., Daulat S.R., Kraft A.S., Cress A.E., Miranti C.K., Mouneimne G., Rogers G.C, & Warfel N.A. (2023). PIM1 phosphorylates ABI2 to enhance actin dynamics and promote tumor invasion. The Journal of Cell Biology, 222(6), e202208136.
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3

Hypoxia signaling protein regulation

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siHIF-1α experiments were performed using a forward transfection protocol (Lipofectamine 3000 Invitrogen). HIF-1α and siRNA was purchased from Santa Cruz Biotechnology (sc-44225 and sc-35316, respectively). PIM1 siRNA sequences were previously reported [58 ]. The following western blot antibodies were used for western blot: actin (BD Biosciences); and HA, HIF-1α (cat #: 3724S), HIF-1α-OH [P564] (cat #: 3434S), p-IRS1 [S1101] (cat #: 2385S), PHD2 (cat #: 4835S), PIM1 (cat #: 4835S), and ubiquitin (cat #: 3933S) (Cell Signaling Technologies). The p-HIF-1α [T455] antibody was generated by Pacific Immunology for western blot and IHC in this manuscript. The following antibodies were also used for IHC: CC3 (cat #: ab4051), CD31 (cat #: ab124432), HIF-1α (cat #: ab16066) and PIM1 (cat #: ab245417)(Abcam).
Casillas A.L., Chauhan S.S., Toth R.K., Sainz A.G., Clements A.N., Jensen C.C., Langlais P.R., Miranti C.K., Cress A.E, & Warfel N.A. (2021). Direct phosphorylation and stabilization of HIF-1α by PIM1 kinase drives angiogenesis in solid tumors. Oncogene, 40(32), 5142-5152.
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4

Quantification of Signaling Proteins

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Total protein for all experiments was collected using RIPA buffer (Sigma, St. Louis, MO) supplemented with 2% Protease Inhibitor Cocktail (Sigma, St. Louis, MO), 2% Phosphatase Inhibitor Cocktail 2 (Sigma, St. Louis, MO), and 2% Phosphatase Inhibitor Cocktail 3 (Sigma, St. Louis, MO). Protein content was assessed from total protein extracts using western immunoblotting with the Criterion apparatus system using 4-15% SDS-polyacrylamide gradient gels (all from Bio-Rad, Hercules, CA) and adjusted to GAPDH (Cat no. AB9484; AbCam, Cambrige, MA). Imaging of western blots was facilitated on the Odyssey infrared imaging system (LiCor, Lincoln, Nebraska). Antibodies against Total Akt (Cat no. 9272), p-S473 Akt (Cat no. 9271), Total IRS1 (Cat no. 2382S), p-S1101 IRS1 (Cat no. 2385), and mTOR (Cat no. 4517) were obtained from Cell Signaling Technology (Danvers, MA) The antibodies for PLIN2 (Cat no. NB110-40877) and PLIN3 (Cat no. NB110-40764) were obtained from Novus Biologicals (Littleton, CO).
Covington J.D., Johannsen D.L., Coen P.M., Burk D.H., Obanda D.N., Ebenezer P.J., Tam C.S., Goodpaster B.H., Ravussin E, & Bajpeyi S. (2017). Intramyocellular Lipid Droplet Size Rather Than Total Lipid Content Is Related to Insulin Sensitivity After 8-Weeks of Overfeeding. Obesity (Silver Spring, Md.), 25(12), 2079-2087.
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5

Antibody Sources for Signaling Pathway Analysis

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Antibodies against the following proteins were obtained from the following sources: alpha-tubulin (ab11304) from Abcam; DEPTOR (NBP149674) from Novus Biologicals; raptor (for IP, A300-553A) from Bethyl laboratories; PLD2 (sc-25513), PPARγ (sc-7196), C/EBPα (sc-61) and aP2 (sc-271529) from Santa Cruz Biotechnology; C/EBPβ (#3087), C/EBPδ (#2318), IRS-1 (#2382), pS636/639-IRS-1 (#2388), pS1101-IRS-1 (#2385), mTOR (#2972), raptor (#2280), Akt (#9272), pS473-Akt (#4051), pT308-Akt (#9275), S6K1 (#9202), pT389-S6K1 (#9234), S6 (#2217), pSer235/236-S6 (#4856), 4EBP1 (#9644), and pT37/46-4EBP1 (#2855) from Cell Signaling Technology. The PLD1 antibody was provided by Dr. Jie Chen from the University of Illinois at Urbana-Champaign16 (link). All secondary antibodies were from Jackson ImmunoResearch Laboratories Inc. [9,10-3H(N)]-oleic acid was from PerkinElmer. C8-PA (830842C) was from Avanti. All other reagents were from Sigma-Aldrich.
Song H.I, & Yoon M.S. (2016). PLD1 regulates adipogenic differentiation through mTOR - IRS-1 phosphorylation at serine 636/639. Scientific Reports, 6, 36968.
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6

Quantification of Signaling Proteins

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Total protein for all experiments was collected using RIPA buffer (Sigma, St. Louis, MO) supplemented with 2% Protease Inhibitor Cocktail (Sigma, St. Louis, MO), 2% Phosphatase Inhibitor Cocktail 2 (Sigma, St. Louis, MO), and 2% Phosphatase Inhibitor Cocktail 3 (Sigma, St. Louis, MO). Protein content was assessed from total protein extracts using western immunoblotting with the Criterion apparatus system using 4-15% SDS-polyacrylamide gradient gels (all from Bio-Rad, Hercules, CA) and adjusted to GAPDH (Cat no. AB9484; AbCam, Cambrige, MA). Imaging of western blots was facilitated on the Odyssey infrared imaging system (LiCor, Lincoln, Nebraska). Antibodies against Total Akt (Cat no. 9272), p-S473 Akt (Cat no. 9271), Total IRS1 (Cat no. 2382S), p-S1101 IRS1 (Cat no. 2385), and mTOR (Cat no. 4517) were obtained from Cell Signaling Technology (Danvers, MA) The antibodies for PLIN2 (Cat no. NB110-40877) and PLIN3 (Cat no. NB110-40764) were obtained from Novus Biologicals (Littleton, CO).
Covington J.D., Johannsen D.L., Coen P.M., Burk D.H., Obanda D.N., Ebenezer P.J., Tam C.S., Goodpaster B.H., Ravussin E, & Bajpeyi S. (2017). Intramyocellular Lipid Droplet Size Rather Than Total Lipid Content Is Related to Insulin Sensitivity After 8-Weeks of Overfeeding. Obesity (Silver Spring, Md.), 25(12), 2079-2087.
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