p irs 1y895 polyclonal rabbit  (Cell Signaling Technology Inc)

Journal: Redox Biology

Article Title: Changes to insulin sensitivity in glucose clearance systems and redox following dietary supplementation with a novel cysteine-rich protein: A pilot randomized controlled trial in humans with type-2 diabetes

doi: 10.1016/j.redox.2023.102918

Figure Lengend Snippet: The effect of 14-weeks of CON, WHEY or KDPWHE treatment in adults with type-2 diabetes mellitus on phosphoprotein concentration within the insulin-receptor signalling pathway upstream of regulatory nodes for GLUT4 translocation within the skeletal muscle in responses to basal (INS -) and insulin stimulation (hyperinsulinemic-isoglycaemic; INS +), with data shown as relative protein abundance at week 0 and 15. (A) Insulin-receptor substrate-1 phosphorylated on Ser312 (IRS-1 Ser312 ). (B) Representative immunofluorescence images of cross-sectional skeletal muscle fibres stained for IRS-1 Ser312 . IRS-1 ser312 was measured as staining intensity in the plasma membrane region (SM ). The small grey scale lines bottom right represent 50 μm. (C) Protein kinase B phosphorylation on Ser437 (Akt Ser437 ) expressed relative to total Akt. (D) Akt substrate 160 kDa phosphorylation on Thr642 (AS160 Thr642 ), (E) p21 activated kinase (PAK) phosphorylation on isoform 1 (PAK Thr423 ) and 2 (PAK Thr402 ). Data are raw unit median, upper and lower quartiles, and range, with individual-participant data points included. Raw unit point and change score mean and SD are in SM Data 1. Outcome statistics are in . CON, non-protein isocaloric control; KDPWHE, keratin-derived protein with whey; WHEY, whey protein isolate.

Article Snippet: The following antibodies were used for visualization of insulin signalling: 1:250 anti-GLUT4 rabbit polyclonal primary antibody (Abcam, catalogue # ab654) [ ], 1:500 mouse polyclonal anti-dystrophin (Sigma-Aldrich, catalogue #D8168), 1:500 rabbit polyclonal anti-p-IRS-1 Ser312 (Invitrogen, catalogue # PA1-1054, p-IRS-1Ser307 in rodent), 1:100 rabbit polyclonal anti-eNOS (Abcam, catalogue # Ab5589), 1:100 rabbit polyclonal anti-p-eNOS Ser1177 (Abcam, catalogue # ab184154) or 1:100 rabbit polyclonal anti-NOX2 (Abcam, catalogue # ab80508) as primary antibodies.

Techniques: Concentration Assay, Translocation Assay, Immunofluorescence, Staining, Membrane, Derivative Assay

Journal: Redox Biology

Article Title: Changes to insulin sensitivity in glucose clearance systems and redox following dietary supplementation with a novel cysteine-rich protein: A pilot randomized controlled trial in humans with type-2 diabetes

doi: 10.1016/j.redox.2023.102918

Figure Lengend Snippet: The effect of 14-weeks of KDPWHE, WHEY, or CON treatment in adults with type-2 diabetes mellitus on skeletal muscle tissue insulin signalling, capillarization, and capillary endothelial nitric oxide synthase activity.

Article Snippet: The following antibodies were used for visualization of insulin signalling: 1:250 anti-GLUT4 rabbit polyclonal primary antibody (Abcam, catalogue # ab654) [ ], 1:500 mouse polyclonal anti-dystrophin (Sigma-Aldrich, catalogue #D8168), 1:500 rabbit polyclonal anti-p-IRS-1 Ser312 (Invitrogen, catalogue # PA1-1054, p-IRS-1Ser307 in rodent), 1:100 rabbit polyclonal anti-eNOS (Abcam, catalogue # Ab5589), 1:100 rabbit polyclonal anti-p-eNOS Ser1177 (Abcam, catalogue # ab184154) or 1:100 rabbit polyclonal anti-NOX2 (Abcam, catalogue # ab80508) as primary antibodies.

Techniques: Activity Assay

Journal: Experimental and Therapeutic Medicine

Article Title: Ursolic acid ameliorates adipose tissue insulin resistance in aged rats via activating the Akt-glucose transporter 4 signaling pathway and inhibiting inflammation

doi: 10.3892/etm.2021.10901

Figure Lengend Snippet: Effect of UA on IRS-1/PI3K/Akt signaling pathway in eWAT of aged rats. (A) Expression of IRS-1, p-IRS-1 (Ser307), p-IRS-1 (Ser307)/IRS-1, Akt, p-Akt (Ser473) and p-Akt (Ser473)/Akt protein levels. (B) PI3K protein levels. Treatment groups were: Young (untreated, n=8), aged (untreated, n=9), treated with 10 mg/kg UA (UA-L, n=8) or 50 mg/kg UA (UA-H, n=9) for 7 weeks. All animals had free access to a standard diet. *P<0.05 compared with aged group. UA-H, high ursolic acid; eWAT, epididymis white adipose tissue; IRS-1, insulin receptor substrate-1; p, phosphorylated; Akt, protein kinase B; PI3K, phosphatidylinositol 3-kinase.

Article Snippet: Membranes were incubated in blocking buffer (5% skimmed milk) for 2 h at room temperature and immunoblotted with the following primary antibodies for 14 h at 4˚C: Rabbit polyclonal anti-IRS-1 antibody (1:1,000; cat. no. TA312859; Origene Technologies, Inc.), rabbit polyclonal anti-phosphorylated(p)-IRS-1 Ser307 antibody (1:1,000; cat. no. TA325573; Origene Technologies, Inc.), rabbit monoclonal anti-Akt antibody (1:1,000; cat. no. ab179463; Abcam), rabbit monoclonal anti-p-Akt Ser473 antibody (1:1,000; cat. no. 4060; Cell Signaling Technology, Inc.), rabbit polyclonal PI3K antibody (1:6,000; cat. no. 2225185; EMD Millipore), rat monoclonal GLUT4 antibody (1:1,000; cat. no. 10613; Santa Cruz Biotechnology, Inc.), rabbit polyclonal NF-κB antibody (1:2,000; cat. no. ab16502; Abcam), rabbit polyclonal IL-1β antibody (1:1,000; cat. no. ab9722; Abcam), rat monoclonal IL-6 antibody (1:1,000; cat. no. ab9324; Abcam), and rabbit polyclonal anti-sodium/potassium-transporting ATPase subunit alpha-1 (ATP-1a1) antibody (1:1,000; cat. no. TA326844; Origene Technologies, Inc.).

Techniques: Expressing