Igf 1 elisa kit

Manufactured by R&D Systems

Sourced in United States

The IGF-1 ELISA kit is a laboratory assay used to quantitatively measure the levels of insulin-like growth factor 1 (IGF-1) in biological samples. The kit utilizes the enzyme-linked immunosorbent assay (ELISA) technique to detect and quantify the target analyte.

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12 protocols using igf 1 elisa kit

Chitosan-Based IGF-1 Encapsulation

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Chitosan (85% deacetylated, medium molecular weight), sodium tripolyphosphate (TPP), acetic acid, phosphate buffered saline (PBS) and silver nitrate were purchased from Sigma-Aldrich (USA). IGF-1 was purchased from Invitrogen (USA), IGF-1 ELISA kit (DY291) was supplied by R&D systems (USA), and LIVE/DEAD cytotoxicity/viability assay (Invitrogen, USA).

Mantripragada V.P, & Jayasuriya A.C. (2014). IGF-1 Release Kinetics from Chitosan Microparticles Fabricated Using Environmentally Benign Conditions. Materials science & engineering. C, Materials for biological applications, 0, 506-516.

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Optimizing Human Hair Protein Expression

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Human hair was obtained by KeraNetics, LLC from World Response Group. VEGF, high binding plates for ELISAs, VEGF ELISA kit, and IGF-1 ELISA kit were obtained from R&D Systems (Minneapolis, MN). IGF-1, bFGF, and bFGF ELISA were obtained from Peprotech (Rocky Hill, NJ). C57BL/6J mice were obtained from Harlan Laboratories (Indianapolis, IN). Ascorbic acid, EDTA, Triton-X100, 28% ammonium hydroxide, Tween 20, and reagents for hematoxylin and eosin (H&E) staining were from Sigma-Aldrich (St. Louis, MO). Horse serum, fetal bovine serum (FBS), high glucose DMEM, low glucose DMEM, DMEM/F12, chick embryo extract, and PBS were obtained from Invitrogen/Life Technologies (Grand Island, NY). Matrigel^™ was obtained from BD Biosciences (San Jose, CA). The Lavacell assay for live cells was from ActiveMotif (Carlsbad, CA). Antibody and immunohistochemical staining kit for blood vessels (von Willebrand factor) was obtained from Millipore (Billerica, MA). Antibody for desmin staining was obtained from Santa Cruz Biotechnology (Santa Cruz, CA). Secondary antibody for HRP staining of desmin was obtained from Vector Laboratories (Burlingame, CA). DC Protein Assay for the modified Lowry method of total protein content was obtained from Bio-Rad (Hercules, CA). Type I collagen was obtained from Worthington Biochemical (Lakewood, NJ).

Tomblyn S., Kneller E.P., Walker S.J., Ellenburg M.D., Kowalczewski C.J., Van Dyke M., Burnett L, & Saul J.M. (2015). Keratin hydrogel carrier system for simultaneous delivery of exogenous growth factors and muscle progenitor cells,,. Journal of biomedical materials research. Part B, Applied biomaterials, 104(5), 864-879.

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Quantifying Metabolic Biomarkers in Serum and Liver

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IGF1 measurements were performed in serum with EDTA using an IGF1 ELISA kit obtained from R&D Systems (MG100). Free fatty acids in serum were assayed using the luminometric Free Fatty Acid Assay Kit from Abnova (KA1667). Triglycerides in liver were assayed using the EnzyChrom Triglyceride Assay Kit from BioAssay Systems (ETGA-200).

Bárcena C., Quirós P.M., Durand S., Mayoral P., Rodríguez F., Caravia X.M., Mariño G., Garabaya C., Fernández-García M.T., Kroemer G., Freije J.M, & López-Otín C. (2018). Methionine Restriction Extends Lifespan in Progeroid Mice and Alters Lipid and Bile Acid Metabolism. Cell Reports, 24(9), 2392-2403.

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hMSCs Isolation and Characterization

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The hMSCs were obtained from the Scientific Research Centre of China-Japan Union Hospital, Jilin University (Changchun, China). A vascular endothelial growth factor (VEGF) enzyme-linked immunosorbent assay (ELISA) kit was obtained from Cosmo Bio Co., Ltd. (Tokyo, Japan). A mouse/rat insulin-like growth factor-1 (IGF-1) ELISA kit was obtained from R&D Systems (Minneapolis, MN, United States). Immunohistochemical antibodies against marker of proliferation Ki-67 (MKI67) and β-catenin (CTNNB) were obtained from Abcam Tec (Shanghai, China). Minoxidil was obtained from Shanghai yuanye Bio-Technology Co., Ltd., (Shanghai, China).

Song D., Pan S., Jin W., Wu R., Zhao T., Jiang J, & Zhu M. (2024). Minoxidil delivered via a stem cell membrane delivery controlled release system promotes hair growth in C57BL/6J mice. Frontiers in Bioengineering and Biotechnology, 11, 1331754.

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Quantifying IGF-1 Secretion Dynamics

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Primary SCs were transfected with IGF-1 siRNA and negative control, miR-129 mimic and control, miR-129 inhibitor and control, respectively. Afterwards, the medium was replaced with FBS-free DMEM for additional incubation. The medium was then taken out and filtered through a 0.22 μm filter (Millipore) to furnish the supernatant. The protein level of IGF-1 in the medium was measured with the IGF-1 ELISA Kit (R&D Systems) according to the manufacturer’s instructions. The data were measured and averaged from three independent cultures, each comprising triplicate wells.

Zhu H., Xue C., Yao M., Wang H., Zhang P., Qian T., Zhou S., Li S., Yu B., Wang Y, & Gu X. (2018). miR-129 controls axonal regeneration via regulating insulin-like growth factor-1 in peripheral nerve injury. Cell Death & Disease, 9(7), 720.

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Quantifying Free IGF-1 in Conditioned Media

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The concentrations of free IGF‐1 within conditioned media were determined using an IGF‐1 ELISA Kit (R&D Systems) under denaturing conditions. To protect the released IGF‐1 and IGF‐binding proteins (IGFBP) from degradation by CG, a protease‐inhibitor cocktail (Sigma) was added to the harvested conditioned media.

Morimoto‐Kamata R, & Yui S. (2017). Insulin‐like growth factor‐1 signaling is responsible for cathepsin G‐induced aggregation of breast cancer MCF‐7 cells. Cancer Science, 108(8), 1574-1583.

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Quantification of IGF-1 in Microglia Supernatants

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Free IGF-1 concentrations in the cell supernatants were determined using the IGF-1 ELISA Kit (R&D System, Wiesbaden, Germany) according to the manufacturer’s instructions. Supernatants were harvested from microglia treated with DMF and MMF for 24 h followed by stimulation with IL-4 or LPS for another 6 h. 50 µL of standard, control, or sample were added per well and incubated for 2 h at room temperature on a horizontal shaker. After washing, IGF-1 conjugate was added to each well and incubated for another 2 h. Following the addition of substrate solution, the enzyme reaction color product was calculated by subtracting wavelengths detection at 570 nm from the readings at 450 nm with Sunrise-Basic Tecan (Tecan, Grödig, Austria).

Kronenberg J., Pars K., Brieskorn M., Prajeeth C.K., Heckers S., Schwenkenbecher P., Skripuletz T., Pul R., Pavlou A, & Stangel M. (2019). Fumaric Acids Directly Influence Gene Expression of Neuroprotective Factors in Rodent Microglia. International Journal of Molecular Sciences, 20(2), 325.

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IGF1 Quantification in Conditioned Medium

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IGF1 in conditioned medium was quantified using IGF1 ELISA kit (R&D Systems). The cell number was counted when the CM was collected, and ELISA data was normalized by 1x10⁶ cells in all cell lines.

Hirakawa T., Yashiro M., Doi Y., Kinoshita H., Morisaki T., Fukuoka T., Hasegawa T., Kimura K., Amano R, & Hirakawa K. (2016). Pancreatic Fibroblasts Stimulate the Motility of Pancreatic Cancer Cells through IGF1/IGF1R Signaling under Hypoxia. PLoS ONE, 11(8), e0159912.

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Quantifying Growth Factors in Bioreactor

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To determine the amount of growth factors in bioreactor supernatant before implantation, culture medium after the 1 week culture period was analyzed using VEGF ELISA kit (#DY293B, R&D Systems), IGF-1 ELISA kit (#DY291, R&D Systems), Adiponectin ELISA kit (#DY1065, R&D Systems), and Thrombospondin ELISA kit (#ab193716, Abcam). The concentration of growth factors was normalized to the total amount of protein measured by the BCA quantification kit.

Guerrero J., Dasen B., Frismantiene A., Pigeot S., Ismail T., Schaefer D.J., Philippova M., Resink T.J., Martin I, & Scherberich A. (2022). T-cadherin Expressing Cells in the Stromal Vascular Fraction of Human Adipose Tissue: Role in Osteogenesis and Angiogenesis. Stem Cells Translational Medicine, 11(2), 213-229.

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Comprehensive Mouse Tissue Analysis

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Prior to sacrifice at 6 or 18 weeks of age, the mice were fasted for 6 h (during dark the phase). The mice were anaesthetized by isoflurane inhalation, which was followed by heart puncture and decapitation. Brains were dissected into different sub-regions, including the hypothalamus and prefrontal cortex and snap frozen in liquid nitrogen. Length and width of the right femur were measured using a digital micro-caliper. Blood was centrifuged at 2,600 g for 10 min. Plasma was subsequently separated and stored at −80°C until analysis of plasma hormones corticosterone (CORT), insulin-like growth factor-1 (IGF1), osteocalcin (OCN), according to the manufacturer’s instructions (EIA Corticosterone kit, Arbor Assays, Ann Arbor, MI, United States; an IGF1 ELISA kit, R&D Systems, Minneapolis, MN, United States; a Mouse OT/BGP ELISA kit, CUSABIO, Houston, TX, United States). Carcasses were dried till constant weight at 103°C [ISO 6496-198 (E)], and this was followed by fat extraction with petroleum ether (Boom BV, Meppel, Netherlands) in a soxhlet apparatus. All organ analyses were performed by a technician blinded to the experimental conditions.

van Heijningen S., Karapetsas G., van der Beek E.M., van Dijk G, & Schipper L. (2022). Early Life Exposure to a Diet With a Supramolecular Lipid Structure Close to That of Mammalian Milk Improves Early Life Growth, Skeletal Development, and Later Life Neurocognitive Function in Individually and Socially Housed Male C57BL/6J Mice. Frontiers in Neuroscience, 16, 838711.

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