Culture supernatants of all probes were prepared in order to quantify the levels of IGFBP3, IGF-1, MMP-1, MMP-3, MMP-9 and TIMP-1. Supernatants were spun at 14.000 g, 4 °C, 10 min, and frozen at -80 °C. Concentrations of all factors were measured using specific ELISA assays (Human IGFBP-3 DuoSet ELISA, Human IGF-I/IGF-1 Quantikine ELISA Kit, Human Total MMP-1 DuoSet ELISA, Human Total MMP-3 DuoSet ELISA, Human MMP-9 DuoSet ELISA, and Human TIMP-1 DuoSet ELISA from RD-Systems). Further, MMP activity in all supernatants was determined using MMP Activity Assay Kit (Abcam). Assay procedures were performed according to user manuals.
Human igf 1 igf 1 quantikine elisa kit
Manufactured by R&D Systems
Sourced in United States
The Human IGF-I/IGF-1 Quantikine ELISA Kit is a quantitative sandwich enzyme immunoassay designed to measure human insulin-like growth factor I (IGF-I) levels in cell culture supernates, serum, and plasma. The kit utilizes a microplate pre-coated with an antibody specific for IGF-I.
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Lab products found in correlation
Mmp activity assay kit, by Abcam (1 mentions)
Human timp 1 duoset elisa, by R&D Systems (1 mentions)
Human mmp 9 duoset elisa, by R&D Systems (1 mentions)
Human total mmp 1 duoset elisa, by R&D Systems (1 mentions)
Human canine porcine insulin quantikine elisa kit, by R&D Systems (1 mentions)
Human igfbp 3 quantikine elisa kit, by R&D Systems (1 mentions)
Softmax pro software 5.4, by Molecular Devices (1 mentions)
Spectramax 190 absorbance plate reader, by Molecular Devices (1 mentions)
Human bdnf duoset elisa kit, by R&D Systems (1 mentions)
5 protocols using human igf 1 igf 1 quantikine elisa kit
1
Quantification of Secretome Proteases
2
Quantifying Porcine IGF-1 and Insulin
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IGF1 and insulin levels were measured in nonfasting pig plasma samples using the human IGF-I/IGF-1 Quantikine ELISA Kit (catalog no. DG100B, R&D Systems) and the Human/Canine/Porcine Insulin Quantikine ELISA Kit (catalog no. DINS00, R&D Systems), following the manufacturer’s protocol.
3
Measurement of IGF-1 and IGFBP-3 in T1DM and Healthy Serum
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IGF-1 and IGFBP-3 levels were measured from T1DM and HC serum samples that were obtained from the MERIT study using the Human IGF-I/IGF-1 Quantikine® ELISA Kit (R & D Systems, Minneapolis, MN, USA) and Human IGFBP-3 Quantikine® ELISA Kit (R & D Systems), respectively, according to the protocol set by the manufacturer. Serum samples underwent a 100-fold dilution in Calibrator Diluent RD5-18. SpectraMax® 190 absorbance plate reader (Molecular Devices, San Jose, CA, USA) and SoftMax® Pro Software 5.4 (Molecular Devices) were used to analyze the data from both the IGF-1 and the IGFBP-3 assays. A standard curve was created by generating a log/log curve fit. Both the IGF-1 and the IGFBP-3 assays were read at 450 nm with correction at 570 nm.
4
Serum IGF-1 and Sclerostin Levels
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The level of IGF-1 was measured from preserved serum by enzyme-linked immunosorbent assay (ELISA) using a Human IGF-I/IGF-1 Quantikine ELISA Kit (R&D Systems, Minneapolis, MN, USA), according to the manufacturer's protocol. The level of serum sclerostin was measured using Human Sclerostin Quantikine ELISA Kit (R&D Systems, Minneapolis, MN, US), as per the manufacturer's protocol. The assays have detection ranges of 0.1–6 ng/mL and 31.3-2,000 pg/mL, respectively.
Demographic information, clinical information, and essential laboratory parameters (age, occupation, body weight, diabetic treatment, HbA1C level, glomerular filtration rate, creatinine level, and alanine transaminase level) were also collected.
Demographic information, clinical information, and essential laboratory parameters (age, occupation, body weight, diabetic treatment, HbA1C level, glomerular filtration rate, creatinine level, and alanine transaminase level) were also collected.
5
Effects of HIIT on Serum Biomarkers
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Blood was drawn from 12‐h fasted participants in the morning into BD Vacutainer 10‐mL increased silica act clot activator, silicone‐coated tube (BD, Franklin Lakes, NJ, USA) before (baseline), and after (6 weeks) HIIT training (Fig. 1 ). Both blood draws (at baseline and after 6 weeks of HIIT) were “resting,” that is, they took place before exercise was performed. Upon collection, blood samples were allowed to clot by leaving them undisturbed at room temperature for ~45 min and then centrifuged at 3488g for 10 min at 4°C. Serum was aliquoted and stored at −80°C prior to use. Serum levels of BDNF, IGF‐1, total CTSB, and pro‐CTSB were measured using human BDNF DuoSet ELISA kit (DY248), human IGF‐I/IGF‐1 Quantikine ELISA kit (DG100), human total Cathepsin B DuoSet ELISA (DY2176) and human pro‐Cathepsin B DuoSet ELISA (DY953) (R&D Systems, Minneapolis, MN) according to manufacturers’ protocols. A standard curve of recombinant BDNF, IGF‐1, total CTSB, and pro‐CTSB was run on each plate. Samples and standards were run in duplicate.
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