Protein factor conjugated PEGylated fibrin gel was prepared as previously described.27 ,29 (link) Briefly, human fibrinogen (Sigma-Aldrich Co.; St. Louis, MO) was reconstituted in Tris-buffered saline (40 mg mL−1, pH 7.8) and reacted with bifunctional SG-PEG-SG (NOF America Corp, Irvine, CA) in 5: 1 PEG: fibrinogen molar ratio with or without the addition of rat SDF-1α (PeproTech Inc.; Rocky Hill, NJ) and human IGF-I (Pepro Tech Inc.; Rocky Hill, NJ). Gel polymerization was induced by the addition of 25 U mL−1 of human thrombin (Sigma). The final concentration of fibrinogen was 10 mg mL−1, PEG 0.5 mg ml−1, SDF-1α 10 μg mL−1, IGF-I 25 μg ml−1. Twenty-four hours post TK-I/R injury, 0.25 mL of empty PEGylated fibrin gel (Peg-Fib; n = 6), SDF-1α conjugated PEGylated fibrin gel (Peg-Fib/SDF-1α; n = 6), SDF-1α and IGF-I conjugated PEGylated fibrin gel (Peg-Fib/SDF-1α/IGF-1; n = 6) was injected into the lateral gastrocnemius (LGAS) muscle of the TK-injured limb. PEG-Fib-containing treatments were injected in liquid form and polymerized in situ. Functional assessments were performed at 14 days of reperfusion.
Human igf1
Manufactured by Thermo Fisher Scientific
Sourced in United States
Human IGF1 is a laboratory reagent used for the quantitative measurement of insulin-like growth factor 1 (IGF-1) levels in human samples. IGF-1 is a hormone that plays a crucial role in growth and development. The product is designed for use in research and diagnostic applications.
Automatically generated - may contain errors
Lab products found in correlation
Human fgf basic, by Thermo Fisher Scientific (3 mentions)
Human scf, by Thermo Fisher Scientific (2 mentions)
Human il 3, by Thermo Fisher Scientific (2 mentions)
Methocult media, by STEMCELL (2 mentions)
Human vegf, by Thermo Fisher Scientific (2 mentions)
Human egf, by Thermo Fisher Scientific (2 mentions)
Human fibrinogen, by Merck Group (1 mentions)
Human thrombin, by Merck Group (1 mentions)
Matrigel basement membrane matrix, by Corning (1 mentions)
Human intesticult organoid growth medium, by STEMCELL (1 mentions)
Eclipse te300, by Nikon (1 mentions)
Protease and phosphatase inhibitor, by Thermo Fisher Scientific (1 mentions)
Igf 1, by Thermo Fisher Scientific (1 mentions)
Nerve growth factor (ngf), by Thermo Fisher Scientific (1 mentions)
Anti tau, by Merck Group (1 mentions)
Leica sp2 confocal microscope, by Leica camera (1 mentions)
Human pdgf bb, by Thermo Fisher Scientific (1 mentions)
Human tgf β1, by Thermo Fisher Scientific (1 mentions)
Human fgf1, by Thermo Fisher Scientific (1 mentions)
Human fgf2, by Thermo Fisher Scientific (1 mentions)
8 protocols using human igf1
1
PEGylated Fibrin Gel Tissue Repair
2
Generation of Organoids with Driver Mutations
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The organoids containing various driver mutations described in Table 1 (NL, A, AT, AKST, AdeCIN TS and AdeCIN TSK) were generated as previously described [8 (link)] using Corning® Matrigel® Basement Membrane Matrix (Corning Inc., Corning, NY, USA) and IntestiCult™ Organoid Growth Medium (Human) (StemCELL Technologies, Vancouver, BC, Canada) supplemented with 100 ng/mL of human IGF-I (Peprotech, Rocky Hill, NJ, USA) and 100 ng/mL of human FGF-basic (Peprotech).
3
Culturing Endothelial Progenitor Cells from PBMNCs
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PBMNCs were harvested at a concentration of 1 × 105 cells/ml and resuspended with 30% FBS/PBS 200 µl. The following recombinant human cytokines were then added to the cells: human SCF (#300-07; PeproTech) at a concentration of 66.7 ng/ml; human VEGF (#100-20; PeproTech) at a concentration of 33.3 ng/ml; human IL3 (#200-03; PeproTech) at a concentration of 13.3 ng/ml; human IGF-1 (#100-11; PeproTech) at a concentration of 33.3 ng/ml; human FGF Basic (#100-18B; PeproTech) at a concentration of 33.3 ng/ml; and, human EGF (#100-15; PeproTech) at a concentration of 33.3 ng/ml. The cell mixture was resuspended with complete MethoCult™ media (#04236; STEMCELL Technologies, Inc., Vancouver, British Columbia, Canada) at a final volume of 2 ml, and then cultured in a 37 °C environment for 14 days [11 (link)]. Endothelial progenitor cell (EPC) colony forming cells (EPC-CFCs) were assessed under phase-contrast light microscopy (Eclipse TE300; Nikon Instruments, Tokyo, Japan). A colony was defined as the presence of at least 50 cells [15 (link)]. All experiments were performed in triplicate. The numbers of colonies of PBMNCs cultured in QQ culture media and in standard culture media were compared.
4
Stimulation of Murine DRG Neurons
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Murine embryonic DRGs were isolated and cultured as described previously [22 (link)]. For IGF1 stimulation experiments, DRG neurons were cultured with neurobasal medium (NB) in the presence of 2 % horse serum, 2 % B27 and 10 ng/ml NGF (PeProTech) for 20 h. Cells were starved for 4 h in NB after three washing steps with NB to minimize NGF levels. DRGs were then stimulated with either 10 ng/ml NGF, 20 ng/ml human IGF1 (PeProTech) or 20 ng/ml human IGF1 and 200 ng/ml mouse IGFBP5 (BP5DU020, GroPep), respectively. Subsequently, cells were washed with phosphate buffer saline and lysed for 10 min at 4 °C with lysis buffer comprising 50 mM Tris–HCl (pH 7.4), 150 mM NaCl, 1 % Triton-X-100, Protease- and Phosphatase inhibitors (Thermo Scientific). Equal volumes of DRG cell lysates were loaded onto 12 % SDS gels and immunoblotted for IGF1R, pIGF1R, AKT, pAKT and calnexin.
5
Murine Motoneuron Culture and IGF1 Stimulation
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Murine embryonic spinal motoneurons were cultured as described [52 (link)]. BDNF, human IGF1 (PeProTech) and mouse IGFBP5 (BP5DU020, GroPep) were used at the concentrations indicated. Cells were initially counted 4 h after plating to obtain the reference value for 100 % survival. After 7 DIV, surviving motoneuron cells were counted again. For IGF1 stimulation experiments, motoneurons were cultured for 5 days with 5 ng/ml BDNF in full medium, and then starved in the same medium without horse serum and BDNF for 12 h. Cells were then stimulated with IGF1. Neurons were fixed with 4 % PFA and stained with anti-Tau (1:1000, Sigma) and anti-IGFBP5 (1:500, Neuromics) antibodies. Cellular morphology was visualized with fluorescence-coupled secondary antibodies (Jackson ImmunoResearch) using a Leica SP2 confocal microscope. Measurements were done with the Leica AS Lite software (Leica).
6
Culturing Mouse Fibroblasts and Pancreatic Stellate Cells
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NIH 3T3 mouse fibroblast cell lines (ATCC CRL-1658) were cultured in 100 × 20 mm and 150 × 25 mm culture dishes (Falcon Corning cat. no. 353003 and 353025) using Dulbecco’s modified Eagle’s medium containing 4.5 g/L D-glucose, L-glutamine, 110 mg/L sodium pyruvate (DMEM, GIBCO 11995-065) with 10% heat inactivated newborn calf serum, NBCS (GIBCO 26010-074). Primary pancreatic stellate cells (PSC) were cultured until passage 8 in DMEM with 10% heat inactivated fetal bovine serum and penicillin-streptomycin. Cultures were allowed to reach 80% confluency before passaging them into new media. Briefly, cells were washed twice with phosphate-buffered saline (PBS) without calcium and magnesium, Wisent 311-010-CL, cells detached using Trypsin-0.25% EDTA (Wisent) and split at a seeding density of 5,000 cells/cm2. Commercial GFs used in this study included the following: human FGF1 (PeproTech, cat. no. 100–17A), human FGF2 (PeproTech, cat. no. 100–18B), human PDGF-BB (PeproTech, cat. no. 100–14B), human IGF1 (PeproTech, cat. no. 100-11), human IGF2 (PeproTech, cat. no. 100-12), human TGF-β1(PeproTech, cat. no. 100–21C).
7
Cell Culture of Glioblastoma and Normal Human Astrocytes
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U87 and U251 cells lines (Chinese Academy of Medical Sciences, Beijing, China) were cultured in DMEM/high glucose supplemented with 10% fetal bovine serum (FBS, Life Technologies, Carlsbad, CA, USA). NHA were purchased from ScienCell Research Laboratories (Carlsbad, CA, USA) and cultured in RPMI-1640 medium with 10% FBS. All the cells were cultured in a humidified incubator (37 °C, 5% CO2). Human IGF1 was purchased from PeproTech (Rocky Hill, NJ).
8
Cytokine-Induced Expansion of PBMNCs
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PBMNCs were harvested at a concentration 1×10 5 cells/ml and resuspended with 30% FBS/PBS 200 µl. The following recombinant human cytokines were then added to the cells: human SCF (#300-07; PeproTech) at a concentration of 66.7 ng/ml; human VEGF (#100 - 20; PeproTech) at a concentration of 33.3 ng/ml; human IL3 (#200-03; PeproTech) at a concentration of 13.3 ng/ml; human IGF-1 (#100 - 11; PeproTech) at a concentration of 33.3 ng/ml; human FGF Basic (#100-18B; PeproTech) at a concentration of 33.3 ng/ml; and, human EGF (#100 - 15; PeproTech) at a concentration of 33.3 ng/ml. The cell mixture was resuspended with complete MethoCult™ media (#04236; STEMCELL Technologies, Inc., Vancouver, British Columbia, Canada) at a nal volume of 2 ml, and then cultured in a 37°C for 14 days. (11) The numbers of colonies of PBMNCs cultured in QQ culture media and in standard culture media were compared.
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