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Recombinant human igf 1

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Recombinant human IGF-1 is a laboratory-produced form of the insulin-like growth factor 1 (IGF-1) protein. IGF-1 is a polypeptide hormone that plays a key role in cell growth and development.

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Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions) Dimethyl sulfoxide (dmso), by Merck Group (2 mentions) Penicillin, by Thermo Fisher Scientific (2 mentions) Streptomycin, by Thermo Fisher Scientific (2 mentions) Hepes buffer, by Lonza (2 mentions) Apo transferrin, by Merck Group (2 mentions) Recombinant human egf, by R&D Systems (2 mentions) Neon transfection system, by Thermo Fisher Scientific (2 mentions) Fetal bovine serum (fbs), by Atlas Biologicals (2 mentions) L glutamine, by Corning (2 mentions) Penicillin streptomycin solution, by Corning (2 mentions) Anti zeb1, by Cell Signaling Technology (1 mentions) Anti phospho p53 ser15, by Cell Signaling Technology (1 mentions) Anti phospho gsk3β, by Cell Signaling Technology (1 mentions) Anti e cadherin, by Cell Signaling Technology (1 mentions) Anti gsk3β, by Cell Signaling Technology (1 mentions) Anti igf ir, by Cell Signaling Technology (1 mentions) Pd98059, by Promega (1 mentions) Anti vimentin, by Cell Signaling Technology (1 mentions) Ly294002, by Merck Group (1 mentions)

31 protocols using recombinant human igf 1

1

IGF-1 Dose-Dependent Metabolic Effects

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Example 5

a) Effects of IGF1 on Metabolic Activity

FIG. 16A and FIG. 16B report the effects of insulin-like growth factor-1 (IGF-1) on glucose utilization in C2C12 cells treated in 2% horse serum. Cells were plated in DMEM plus 4.5 g/L of glucose and 10% fetal bovine serum (FBS) at Day −2 (d-2). At Day 0 (d0) media was refreshed with DMEM plus 1 g/L glucose and 2% horse serum. At Day 5 (d5) various treatments were added with glucose utilization determined at Day 6 (d6). FIG. 16A reports the dose-response relationship between recombinant human IGF-1 treatment (x-axis) and glucose utilization, revealing an EC50 of 17.43 ng/mL. Recombinant human IGF1 was purchased from R&D Systems (Cat. 291-G1). FIG. 16B reports the dose-response relationship between PPF1 treatment and glucose utilization, revealing an EC50 of 2.9 mg/ml containing 0.87 ng/mL IGF1. IGF-1 is known to have an impact on metabolism of myotubes, and its presence in PPF1 has been calculated as approximately 14.88 ng/mL. However, the data presented here reveals that PPF1 has 20× more efficacy than IGF-1 alone, thus the presence of IGF-1 alone cannot explain the enhanced efficacy observed with PPF1. Thus, other factors must necessarily be involved in the effects of PPF1.

US11439687B2. Blood plasma fractions for use in muscle regeneration (2022-09-13). Alkahest, Inc. [US]. Inventors: Viktoria Kheifets [US], Benson Lu [US], Annette Tennstaedt [US].
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2

Investigating IGF-I Signaling Pathways

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Recombinant human IGF-I was purchased from R&D System (Wiesbaden, Germany). The dual IGF-IR/IR inhibitor OSI-906 was purchased from SelleckBio (USA). Specific PI3K/Akt inhibitor LY294002 was purchased from Sigma (St. Louis, MO), and specific ERK1/2 inhibitor PD98059 was purchased from Promega (Madison, WI). Proteasome inhibitor bortezomib (PS-341) was purchased from Millenium Pharmaceuticals Inc (Cambridge, MA, USA). Anti-E-cadherin, anti-Vimentin, anti-ZEB1, anti-IGF-IR, anti-phospho-IGF-IR (Tyr1131), anti-phospho-GSK-3β, anti-GSK-3β and anti-phospho-P53 (Ser15) were purchased from Cell Signaling Technology (Beverly, MA). Anti-Snail and anti-Twist2 were purchased from Abcam (Cambridge, MA). All the other antibodies were purchased from Santa Cruz Biotechnology (USA).
Li H., Xu L., Li C., Zhao L., Ma Y., Zheng H., Li Z., Zhang Y., Wang R., Liu Y, & Qu X. (2014). Ubiquitin ligase Cbl-b represses IGF-I-induced epithelial mesenchymal transition via ZEB2 and microRNA-200c regulation in gastric cancer cells. Molecular Cancer, 13, 136.
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3

Pharmacological Modulation of IGF-1R in NPC

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Five human NPC cell lines (CNE-1, CNE-2, SUNE-1, 5-8F, and 6-10B) were kindly provided by Prof. Yunfei Xia (Sun Yat-Sen University Cancer Center, Guangzhou, China). NPC cell lines were maintained in RPMI-1640 supplemented with 10% fetal bovine serum (FBS), 100 units/ml penicillin, 100 mg/ml streptomycin, and 2 mM of glutamine and cultured at 37°C with a humidified 5% CO2.
The linsitinib (IGF-1R inhibitor) was obtained from Selleckchem (Houston, TX, USA) and dissolved in DMSO (Sigma-Aldrich) at a concentration of 10 mM. 0.1% DMSO was used to be a control treatment of 10 μM linsitinib. Recombinant human IGF-I was purchased from R&D System (Wiesbaden, Germany).
Wang Z., Liu G., Mao J., Xie M., Zhao M., Guo X., Liang S., Li H., Li X, & Wang R. (2019). IGF-1R Inhibition Suppresses Cell Proliferation and Increases Radiosensitivity in Nasopharyngeal Carcinoma Cells. Mediators of Inflammation, 2019, 5497467.
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4

Promastigote-Induced Foot Lesion Measurement

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Stationary-phase promastigotes (1 × 106) that had been preincubated with or without 50 ng/mL of recombinant human IGF-I (R&D Systems, USA) for 5 min and then washed were injected into the footpads of BALB/c and C57BL/6 mice. In the opposite footpad, we injected RPMI 1640 medium as a control. For six weeks, we measured the thickness of the foot to indicate the growth of the lesions using a micrometer (Mitsutoyo, Japan), and the difference between the infected and noninfected footpads in millimeters was considered the lesion size.
Reis L.C., Ramos-Sanchez E.M., Petitto-Assis F., Nerland A.H., Hernandez-Valladares M., Selheim F., Floeter-Winter L.M, & Goto H. (2018). Insulin-Like Growth Factor-I as an Effector Element of the Cytokine IL-4 in the Development of a Leishmania major Infection. Mediators of Inflammation, 2018, 9787128.
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5

Quantifying IGF Receptor Signaling in Lung Cancer Cells

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A panel of human lung cancer derived cell lines (ATCC, Table 1) were assessed by ELISA for the presence of PAPP-A in the culture medium. Human alveolar adenocarcinoma cells (A549, ATCC) [50 (link)] were selected and thereafter maintained in DMEM containing 10% FBS, 2 mM glutamine, penicillin and streptomycin. The cells were allowed to reach confluence in a 96-well plate, washed three times and switched to DMEM containing antibiotics and 0.1% bovine serum albumin for 16 hours to allow PAPP-A secretion and accumulation in the medium. Varying doses of an irrelevant control IgG2a antibody (BioXCell) or PAPP-A mAb 1/41 were added 1 h prior to the addition of recombinant human IGF-I (50 ng/ml, R&D Systems), or IGF-I and human recombinant IGFBP-4 (50/500 ng/ml, R&D Systems) pre-incubated for 30 minutes to allow the IGF/IGFBP-4 complex to form. Fifteen minutes after IGF stimulation, conditioned media were collected and cells fixed and permeabilized for In-Cell Western analysis (LI-COR). An anti-phospho-AKT antibody (Novus Biologicals) was used to assess IGF receptor signaling using the LI-COR Odyssey system. Levels of PAPP-A were determined by ELISA (AL-101, Ansh Labs).
Mikkelsen J.H., Resch Z.T., Kalra B., Savjani G., Kumar A., Conover C.A, & Oxvig C. (2014). Indirect targeting of IGF receptor signaling in vivo by substrate-selective inhibition of PAPP-A proteolytic activity. Oncotarget, 5(4), 1014-1025.
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6

Inhibiting IGF-IR and Src Signaling

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Recombinant human IGF-I was purchased from R&D System (Wiesbaden, Germany). OSI-906, the dual IGF-IR/IR inhibitor, was purchased from SelleckBio (Houston, TX, USA). PP2 was purchased from Sigma-Aldrich (St. Louis, MO, USA). Antibodies to E-cadherin, vimentin, IGF-IR, phospho-IGF-IR (Tyr1131), Src, and phospho-Src (Y416) were purchased from Cell Signaling Technology Inc. (Beverly, MA, USA). Anti-actin and secondary goat anti-rabbit and goat anti-mouse antibodies were purchased from Santa Cruz Biotechnology Inc. (Santa Cruz, CA, USA).
Wang R., Li H., Guo X., Wang Z., Liang S, & Dang C. (2016). IGF-I Induces Epithelial-to-Mesenchymal Transition via the IGF-IR–Src–MicroRNA-30a–E-Cadherin Pathway in Nasopharyngeal Carcinoma Cells. Oncology Research, 24(4), 225-231.
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7

Modulators of Estrogen and IGF Signaling

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Penicillin, streptomycin, charcoal-stripped fetal bovine serum (CS-FBS), and fetal bovine serum (FBS), were purchased from Invitrogen (Camarillo, CA, USA). The agonist G-1 of the G protein-coupled estrogen receptor-1 (GPER-1) and the ER antagonist ICI 182,780 were from TOCRIS (Bristol, UK); E2, 5α-Dihydrotestosterone (DHT), or E2-BSA-FITC, 4-hydroxytamoxifen (4-OHT) were obtained from Sigma (St. Louis, MO, USA). The recombinant human IGF-I was from R&D Systems (Minneapolis, MN, USA) and the recombinant human IFG-II was from PerproTech (Rocky Hill, NJ, USA). The selective inhibitors of the Src family of protein tyrosine kinases (PP2) and IGF-1R (AG1024) were from EMD Millipore (Calbiochem, Temecula, CA). The NF-κB activation inhibitor II was purchased from EMD Millipore. Unless otherwise indicated, all other drugs were obtained from Sigma-Aldrich (St. Louis, MO, USA).
Kim H., Datta A., Talwar S., Saleem S.N., Mondal D, & Abdel-Mageed A.B. (2016). Estradiol-ERβ2 signaling axis confers growth and migration of CRPC cells through TMPRSS2-ETV5 gene fusion. Oncotarget, 8(38), 62820-62833.
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8

Mosquito Rearing and Blood Feeding

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Immortalized, ASE cell lines were maintained as previously described [9] (link). For in vivo studies, A. stephensi Liston (Indian wild-type strain) were reared and maintained at 27°C and 75% humidity. All mosquito rearing and feeding protocols were approved and in accordance with regulatory guidelines and standards set by the Institutional Animal Care and Use Committee of the University of California, Davis. For most experimental treatments, laboratory-reared 3–5 day old female mosquitoes were maintained on water for 24–48 h and then allowed to feed for 30 min on reconstituted human blood meals (healthy male adult, blood type O+) provided through a Hemotek Insect Feeding System (Discovery Workshops, Accrington, UK). Reconstituted blood meals contained washed human red blood cells (RBCs) and saline (10 mM, NaHCO3, 15 mM NaCl, pH 7.0) as 50% RBCs and 50% saline with or without Recombinant human IGF1. Human RBCs were purchased from Interstate Blood Bank (Memphis, TN, USA). Recombinant human IGF1 was purchased from R&D Systems (Minneapolis, MN, USA).
Drexler A.L., Pietri J.E., Pakpour N., Hauck E., Wang B., Glennon E.K., Georgis M., Riehle M.A, & Luckhart S. (2014). Human IGF1 Regulates Midgut Oxidative Stress and Epithelial Homeostasis to Balance Lifespan and Plasmodium falciparum resistance in Anopheles stephensi. PLoS Pathogens, 10(6), e1004231.
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9

Bovine Ovarian Cell Culture Protocol

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There were no live animals used in this study, so no ethical approval was required. Bovine ovaries were collected at an abattoir where humane slaughter practices were followed, according to USDA guidelines. The hormones and reagents used in cell culture were: ovine follicle-stimulating hormone (FSH; NIDDK-oFSH-20; activity: 175 × NIH-FSH-S1 U/mg) from the National Hormone and Pituitary Program (Torrance, CA, USA), recombinant human IGF1 from R&D Systems (Minneapolis, MN, USA); testosterone from Steraloids (Wilton, NH, USA); and fetal calf serum (FCS) from Atlanta Biologicals (Atlanta, GA, USA); ENNA, Dulbecco’s modified Eagle’s medium (DMEM), Ham’s nutrient mixture F-12 (F12), collagenase, DNase, gentamycin, glutamine, and sodium bicarbonate from Sigma-Aldrich, Inc. (St. Louis, MO, USA).
Chiminelli I., Spicer L.J., Maylem E.R, & Caloni F. (2022). In Vitro Effects of Enniatin A on Steroidogenesis and Proliferation of Bovine Granulosa Cells. Toxins, 14(10), 714.
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10

IGFBP7 Modulates Myeloma Cell Growth

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Cell Counting Kit-8 (Sigma) was used to analyze the impact of recombinant human IGFBP7 (rhIGFBP7; Peprotech) on myeloma cell growth following the manufacturer’s directions. In brief, 2 × 104 cells per well were seeded in flat-bottomed 96-well plates (Iwaki) in the presence of rhIGFBP7 (0–10 μg/ml). Neutralization experiments with recombinant human IGF-1 (10 or 100 ng/ml) (R&D Systems), insulin (10 or 100 ng/ml) (Roche Diagnostics) and IGFBP7 (1 or 10 μg/ml) were performed in serum free Syn-H medium (ABCell-Bio) according to Sprynski et al. [6 (link)]
Bolomsky A., Hose D., Schreder M., Seckinger A., Lipp S., Klein B., Heintel D., Ludwig H, & Zojer N. (2015). Insulin like growth factor binding protein 7 (IGFBP7) expression is linked to poor prognosis but may protect from bone disease in multiple myeloma. Journal of Hematology & Oncology, 8, 10.
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