Fgf21

Manufactured by Merck Group

Sourced in United States, Germany

FGF21 is a recombinant human fibroblast growth factor 21 protein. It is a member of the fibroblast growth factor family and plays a role in glucose and lipid metabolism.

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Lab products found in correlation

Igf 1, by R&D Systems (7 mentions) Insulin, by ALPCO (6 mentions) Adiponectin, by R&D Systems (5 mentions) Leptin, by R&D Systems (4 mentions) Fgf21, by R&D Systems (3 mentions) Glp 1, by Phoenix Pharmaceuticals (3 mentions) Insulin, by Merck Group (3 mentions) Adiponectin, by Merck Group (2 mentions) Leptin, by Merck Group (2 mentions) Mbnmag 41k, by Merck Group (2 mentions) Luminex 200 system, by Merck Group (2 mentions) Spectramax m5 microplate reader, by Molecular Devices (2 mentions) Freestyle glucometer and test strips, by Abbott (2 mentions) Blood glucose meter, by Bayer (1 mentions) Ang 2, by R&D Systems (1 mentions) Fgf19, by R&D Systems (1 mentions) Vascular endothelial growth factor (vegf), by R&D Systems (1 mentions) Vegf a, by Merck Group (1 mentions) Fibroblast growth factor 2 (fgf2), by Merck Group (1 mentions) Chemerin, by Abcam (1 mentions)

26 protocols using fgf21

Plasma Biomarker Quantification Protocol

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After blood collection, glucose was determined using a blood glucose meter (Bayer Consumer Care AG, Basel, Switzerland). Then, plasma was separated by centrifugation and stored at –20 °C until assayed. Plasma total cholesterol, triglycerides and P were measured by spectrophotometry (BioSystems SA, Barcelona, Spain). ELISA tests were used to quantify plasma FGF21 (EMD Millipore Corporation, St. Charles, MO, USA), leptin and adiponectin (EMD Millipore Corporation, St. Charles, MO, USA).

Pineda C., Rios R., Raya A.I., Rodriguez M., Aguilera-Tejero E, & Lopez I. (2018). Hypocaloric Diet Prevents the Decrease in FGF21 Elicited by High Phosphorus Intake. Nutrients, 10(10), 1496.

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Measurement of Serum Angiogenic Factors

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Serum VEGF, ANG2, FGF19, and FGF21 levels were evaluated using commercial enzyme-linked immunosorbent assays (VEGF, ANG2, and FGF19: R&D Systems, Minneapolis, MN, USA; FGF21: Merck Millipore, Darmstadt, Germany) according to the manufacturers’ protocols [14 (link)].

Shigesawa T., Suda G., Kimura M., Maehara O., Tokuchi Y., Kubo A., Yamada R., Furuya K., Baba M., Kitagataya T., Suzuki K., Ohara M., Kawagishi N., Nakai M., Sho T., Natsuizaka M., Morikawa K., Ogawa K, & Sakamoto N. (2021). Baseline serum angiopoietin-2 and VEGF levels predict the deterioration of the liver functional reserve during lenvatinib treatment for hepatocellular carcinoma. PLoS ONE, 16(3), e0247728.

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Biomarker Assessment of Metabolic and Vascular Health

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Study subjects venous blood samples were collected after overnight fasting, centrifuged, and stored at −80 °C. TRX1 (Sigma-Aldrich Chemie GmbH, St. Louis, MO, USA, RAB1756-1KT), MPO (Cayman Chemical, Ann Arbor, MI, USA, 501410), Chemerin (Abcam, Cambridge, UK, ab155430), GDF-15 (Sigma-Aldrich Chemie GmbH, RAB0204-1KT), Ang-2 (Sigma-Aldrich Chemie GmbH, RAB0016-1KT), VEGF-A (Sig-ma-Aldrich Chemie GmbH, RAB0507-1KT), FGF-2 (Sigma-Aldrich Chemie GmbH, RAB0182-1KT), FGF-21 (Merck KGaA, Darmstadt, Germany, EZHFGF21-19K), MMP-1, -3, -9 (Merck KGaA, QIA55-1EA; RayBiotech Life, Inc., Peachtree Corners, GA, USA, ELH-MMP3-1-RB; RayBiotech Life, Inc., ELH-MMP9-001) and CRP (Sigma-Aldrich Chemie GmbH, RAB0096-1KT) were measured in plasma by ELISA method using TECAN Infinite 200 PRO multimode reader (Tecan Group, Ltd., Mannedorf, Switzerland). Concentrations of lipids, glucose, and other routine blood biomarkers were analyzed by standard methods.

Tretjakovs P., Lurins J., Svirskis S., Gersone G., Lurina D., Rozenberga U., Blumfelds L., Bahs G., Lejnieks A, & Mackevics V. (2021). Thioredoxin-1 and Correlations of the Plasma Cytokines Regarding Aortic Valve Stenosis Severity. Biomedicines, 9(8), 1041.

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Metabolic Markers Determination Protocol

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Commercially available spectrophotometric kits were used for triglyceride determination (Biosystems, Barcelona, Spain), and ELISA kits for insulin (Millipore, Darmstadt, Germany), leptin (Sigma Aldrich, St Louis, MO, USA), adiponectin (Millipore, Darmstadt, Germany), and FGF-21 (Merck KGaA, Darmstadt, Germany) measurements in serum samples.
The homeostatic model assessment for insulin resistance (HOMA-IR) was calculated from basal insulin and glucose values using Matthews' formula. 23 HOMA-IR ¼ ½fasting glucose ðmmol L À1 Þ Â fasting insulin ðmU L À1 Þ=22:5:
The triglyceride and glucose index (TyG) was calculated as an additional marker of insulin resistance using the formula proposed by Simental-Mendía et al. 24 TyG ¼ Ln ðTG ½mg dL À1 Â glucose ½mg dL À1 =2Þ

Arellano-García L., Macarulla M.T., Cuevas-Sierra A., Martínez J.A., Portillo M.P, & Milton-Laskibar I. (2023). Lactobacillus rhamnosus GG administration partially prevents diet-induced insulin resistance in rats: a comparison with its heat-inactivated parabiotic. Food & function, 14(19).

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FGF21-Induced Signaling in HEK293T Cells

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HEK293T cells were cultured in 75 cm² cell culture flasks, and the cells were then treated with the indicated concentrations of MG when they reached 40–50% confluency. The cells were subsequently lysed following the above-described procedures, and 10 μg whole cell lysate samples were used for Western blot analysis (see Supplementary Materials).
Phospho-FRS2-α (Tyr196) antibody (Cell Signaling # 3864, with a 1:1000 dilution), Phospho-FGFR (Tyr653/654) antibody (Cell Signaling #3471, with a 1:1000 dilution), EGFR 1005 antibody (Santa Cruz Biotechnology, sc-03, with a 1:10000 dilution), IGF2R H-20 antibody (Santa Cruz Biotechnology, sc-14408, with a dilution ratio of 1:1000), and FRS2 H-91 antibody (Santa Cruz Biotechnology, sc-14408, with a dilution ratio of 1:1000) were employed as primary antibodies. Horseradish peroxidase-conjugated anti-rabbit IgG and Alexa Fluor® 647 donkey anti-goat IgG (H+L) were used as secondary antibodies. Membranes were also probed with anti-actin antibody (Cell Signaling #4967, at a dilution ratio of 1:10000) to confirm equal protein loading. Cells were treated with 100 ng/mL FGF21 (Sigma-Aldrich) for 20 min before harvesting for Western blot analysis of phospho-FRS2-α and phospho-FGFR.

Miao W., Xiao Y., Guo L., Jiang X., Huang M, & Wang Y. (2016). A High-throughput Targeted Proteomic Approach for Comprehensive Profiling of Methylglyoxal-induced Perturbations of the Human Kinome. Analytical chemistry, 88(19), 9773-9779.

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FGF21 Modulates TNFα-Induced Muscle Atrophy

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The murine myoblast cell line C2C12 (5 × 10⁵ cells/mL) was grown at 37 °C in 5% CO₂ in DMEM (Life Technologies, Grand Island, NY, USA) containing 10% FBS (Life Technologies) and 1% penicillin-streptomycin (Life Technologies). At a confluence of 95–100%, the medium was replaced with differentiation medium [DMEM plus 2% horse serum (Life Technologies)], which was changed after 2 days. To examine the effects of FGF21 on TNFα-induced muscle atrophy, differentiated myotubes were treated for 24 h with 100 ng/mL of TNFα (Pepro Tech), rmFGF21 (Creative Biomart, Shirley, NY, USA), or both drugs in combination. To examine the link between the effects of FGF21 on TNFα-induced muscle atrophy and AMPK, C2C12 myotubes were treated with rmFGF21, 20 μM compound C (AMPK inhibitor, Sigma), and/or 0.5 mM 5-aminoimidazole-4-carboxamide ribonucleotide (AICAR, AMPK activator, Sigma).

Kim C.S., Joe Y., Choi H.S., Back S.H., Park J.W., Chung H.T., Roh E., Kim M.S., Ha T.Y, & Yu R. (2019). Deficiency of fibroblast growth factor 21 aggravates obesity-induced atrophic responses in skeletal muscle. Journal of Inflammation (London, England), 16, 17.

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Measuring Plasma Peptide Levels

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Plasma concentrations of the peptides were measured using EIA kits: ghrelin, CCK, GLP-1 (Phoenix Pharmaceuticals, Inc., USA), and FGF-21 (Millipore Corporation, USA). The sensitivity of the methods are provided by kit suppliers and are as follows: ghrelin – 0.08 ng/ml (14% intra- assay and 5% interassay variability),CCK – 0.06 ng/ml (5% intra- assay and 9% interassay variability), GLP-1 – 0.18 ng/ml (14% intra- assay and 5% interassay variability), FGF-21 – 0.016 ng/ml (5.8% intra- assay and 9% interassay variability).

Skoczeń S., Rej M., Kwiecińska K., Pietrys D., Tomasik P.J., Wójcik M., Strojny W., Dłużniewska A., Klimasz K., Fijorek K., Korostyński M., Piechota M, & Balwierz W. (2020). Gastrointestinal peptides in children before and after hematopoietic stem cell transplantation. BMC Cancer, 20, 306.

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Hormone Concentration Measurement Protocols

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We assessed the concentrations of various hormones by the EIA method: CCK, ghrelin, GLP-1 (Phoenix Pharmaceuticals, Inc., Burlingame, CA, USA) and FGF-21 (Millipore Corporation, Burlington, MA, USA). Plasma samples for adipokine and soluble leptin receptor analyses were stored at −80 °C until the time of the measurement. Plasma concentrations of the peptides were measured using the following assays: enzyme immunoassay (EIA) (Phoenix Pharmaceuticals, Burlingame, CA, USA)—adiponectin, apelin and visfatin; enzyme-amplified sensitivity immunoassay (Biosource; Nivelles, Belgium)—leptin; (EIA) (BioVendor Research and Diagnostic Products, Brno, Czech Republic)—leptin receptor and resistin.

Czogała W., Strojny W., Schab M., Grabowska A., Miklusiak K., Kowalczyk W., Łazarczyk A., Tomasik P, & Skoczeń S. (2021). FTO and PLAG1 Genes Expression and FTO Methylation Predict Changes in Circulating Levels of Adipokines and Gastrointestinal Peptides in Children. Nutrients, 13(10), 3585.

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Glucose Uptake Assay Protocol

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2-Deoxy-d-[2,6-³H]-glucose was purchased from PerkinElmer NEN radiochemicals. Insulin, dexamethasone, isobutylmethylxanthine (IBMX), cytochalasin B, glucose, 2-deoxyglucose, 5-Aza, ECCG, and TF-3 were purchased from Sigma-Aldrich. Recombinant TNF-α and Fgf21 were purchased from Millipore. RG-108 was purchased from Stemgent.

You D., Nilsson E., Tenen D.E., Lyubetskaya A., Lo J.C., Jiang R., Deng J., Dawes B.A., Vaag A., Ling C., Rosen E.D, & Kang S. (2017). Dnmt3a is an epigenetic mediator of adipose insulin resistance. eLife, 6, e30766.

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Insulin and FGF21 ELISA Analysis

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Serum Insulin and FGF21 were analyzed via ELISA (Insulin, Millipore, Billerica, MA; FGF21 – R&D Systems, Minneapolis, MN).

Forney L.A., Wanders D., Stone K.P., Pierse A, & Gettys T.W. (2017). CONCENTRATION-DEPENDENT LINKAGE OF DIETARY METHIONINE RESTRICTION TO THE COMPONENTS OF ITS METABOLIC PHENOTYPE. Obesity (Silver Spring, Md.), 25(4), 730-738.

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