Eglp 35k

Manufactured by Merck Group

Sourced in United States, Germany, Japan, United Kingdom

The EGLP-35K is a laboratory equipment product manufactured by the Merck Group. It is designed for general laboratory use. The core function of the EGLP-35K is to perform laboratory tasks, but a detailed description is not available while maintaining an unbiased and factual approach.

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Close spelling variants of this product

GLP-1 (Active) ELISA Kit (EGLP-35K, (6 citations)

33 protocols using eglp 35k

Oral Glucose Tolerance Tests for GLP-1 and Insulin Sensitivity

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Oral-glucose-tolerance tests (OGTT, 75 g, 2 hr) were performed in the morning after an overnight fast. Subjects habitually consumed ≥150 g/day carbohydrate prior to the OGTT and were instructed to refrain from exercise for ≥48 h. Venous blood was collected into EDTA containing tubes before and every 30 min after the oral glucose load. Samples to be analyzed for GLP-1 were collected into tubes that also contained dipeptidyl peptidase-4 inhibitor (DPP4, Millipore Corp., Saint Charles, MO) to prevent the degradation of active GLP-1. Plasma was isolated and frozen at −80º C for later analysis.
Plasma glucose was measured by using the glucose oxidase method (Stat Plus, YSI Corp., Yellow Springs, OH). Insulin was measured with a double-antibody radioimmunoassay (17 ). Total GIP and active GLP-1 (7–36 and 7–37 amides) were measured with ELISA assays (EZHGIP-54K and EGLP-35K, respectively, Millipore Corp., Saint Charles, MO). All plasma analytes were measured by personnel who were blinded to subject identities and study groups. Positive incremental area under the curve (AUC) was calculated by using the trapezoidal method (3 (link)). Insulin sensitivity index (ISI) was calculated from OGTT glucose and insulin data (15 (link)). Homeostasis model assessment of insulin resistance (HOMA-IR) was calculated from fasting glucose and insulin (16 (link)).

Weiss E.P., Royer N.K., Fisher J.S., Holloszy J.O, & Fontana L. (2014). Postprandial Plasma Incretin Hormones in Exercise-Trained versus Untrained Subjects. Medicine and science in sports and exercise, 46(6), 1098-1103.

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Oral d-Allulose Effects on Gut Hormones

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The blood samples were collected from the portal vein of male mice fasted overnight (18:00 to next 10:00) under isoflurane anesthesia at 0, 1, 2, and 3 h after p.o. administration of d-allulose (0.3 and 1 g kg⁻¹, 10 ml kg⁻¹) or saline (10 ml kg⁻¹), respectively. The sampling syringe contained heparin (final concentration; 50 IU ml⁻¹), aprotinin (final concentration; 500 kIU ml⁻¹), and DPP-IV inhibitor vildagliptin (final concentration; 10 µM). Plasma was collected after centrifugation (3000 × g, 10 min at 4 °C) and stored at –80 °C until assay. Active GLP-1, total GIP, PYY, and CCK levels were measured using GLP-1 (Active) ELISA (EGLP-35K; Millipore), Rat/Mouse GIP (total) ELISA (EZRMGIP-55K; Millipore), Mouse/Rat PYY EIA (YK081, Yanaihara Institute, Inc.), and Human/Rat/Mouse Cholecystokinin Octapeptide (26-33, non-sulfated) EIA kits (Phoenix Pharmaceuticals, Inc.), respectively.

Iwasaki Y., Sendo M., Dezaki K., Hira T., Sato T., Nakata M., Goswami C., Aoki R., Arai T., Kumari P., Hayakawa M., Masuda C., Okada T., Hara H., Drucker D.J., Yamada Y., Tokuda M, & Yada T. (2018). GLP-1 release and vagal afferent activation mediate the beneficial metabolic and chronotherapeutic effects of D-allulose. Nature Communications, 9, 113.

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Quantifying Serum FGF, GLP-1 Levels

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Serum levels of fibroblast growth factor 19 (FGF19) (catalog #DF1900; R&D Systems, Minneapolis, MN), FGF21 (catalog #DF2100; R&D Systems), and glucagon‐like peptide 1 (GLP‐1) (catalog #EGLP‐35K; Millipore, Burlington, MS) were conducted per the manufacturers’ protocols.

Yang Z., Kusumanchi P., Ross R.A., Heathers L., Chandler K., Oshodi A., Thoudam T., Li F., Wang L, & Liangpunsakul S. (2019). Serum Metabolomic Profiling Identifies Key Metabolic Signatures Associated With Pathogenesis of Alcoholic Liver Disease in Humans. Hepatology Communications, 3(4), 542-557.

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Plasma Glucose, Insulin, and Incretins Dynamics

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On days 0 and 14 and throughout the 3-h meal challenge on day 14, blood will be collected for measures of plasma glucose, insulin, and incretins. Glucose will be assessed by clinical assay (Pointe Scientific). Plasma insulin (ALPCO, #80-INSHU-E10.1) as well as the gut incretins gastric inhibitory peptide (GIP, #EZHGIP-54K; Millipore) and glucagon-like-peptide-1 (GLP-1; #EGLP-35K; Millipore) will be assessed using separate ELISA kits in accordance with the manufacturer's instructions.

Quarles W.R., Pokala A., Shaw E.L., Ortega-Anaya J., Hillmann L., Jimenez-Flores R, & Bruno R.S. (2020). Alleviation of Metabolic Endotoxemia by Milk Fat Globule Membrane: Rationale, Design, and Methods of a Double-Blind, Randomized, Controlled, Crossover Dietary Intervention in Adults with Metabolic Syndrome. Current Developments in Nutrition, 4(9), nzaa130.

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In Vivo and In Vitro GLP-1 Secretion Assay

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For in vivo GLP-1 secretion, mice were fasted for 6 h, gavaged with 25 mg per kg (body weight) sitagliptin (Sigma) 1 h before a glucose bolus (2 g per kg body weight). Fifteen minutes after glucose gavage, mice were killed by CO₂ asphyxiation and portal vein blood or peripheral blood was collected. Serum GLP-1 was measured as described below.
For in vitro GLP-1 secretion, intestinal organoids were isolated from Myc^fl/+ and Myc^ΔIE/+ mice fed a HFD for 2 weeks and cultured for 7 d. On day 7, after 30 min glucose deprivation in DMEM no-glucose medium, a 1-h GLP-1 secretion test in response to DMEM without glucose and to DMEM with glucose (5.5 mmol l⁻¹) was performed at 37 °C in DMEM plus 1% dipeptidyl peptidase-IV inhibitor (Millipore). Finally, the medium was collected and centrifuged for 5 min at 4 °C at 13,000g. For the MYC inhibitor experiment, organoids were pretreated with DMSO or 40 μM 10058-F4 24 h before the GLP-1 secretion test. Medium GLP-1 was measured as described below. The protein content of the remaining organoids was assessed by the BCA protein assay kit (Pierce Chemical).
Total GLP-1 and active GLP-1 were measured by ELISA kits EZGLP1T-36K and EGLP-35K (Millipore), respectively. Medium GLP-1 levels were normalized to the total quantity of cellular protein.

Luo Y., Yang S., Wu X., Takahashi S., Sun L., Cai J., Krausz K.W., Guo X., Dias H.B., Gavrilova O., Xie C., Jiang C., Liu W, & Gonzalez F.J. (2021). Intestinal MYC modulates obesity-related metabolic dysfunction. Nature metabolism, 3(7), 923-939.

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Analytical Techniques for Metabolic Hormones

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Blood samples were collected from the tail vein of animals into ice-chilled heparin-coated microcentrifuge tubes. Blood glucose was measured using a portable Ascencia meter (Bayer Healthcare, Newbury, Berkshire, UK). For plasma insulin and glucagon, blood was collected in chilled fluoride/heparin-coated tubes (Sarstedt, Numbrecht, Germany) and centrifuged using a Beckman microcentrifuge (Beckman Instruments, Galway, Ireland) for 10 minutes at 12,000 rpm. Plasma was extracted and stored at - 20°C. For hormone determination from tissues, samples underwent acid-ethanol extraction (HCl: 1.5% v/v, ethanol: 75% v/v, H₂O:23.5% v/v). Insulin concentrations were subsequently assessed by an in-house radioimmunoassay [28 (link)]. Plasma glucagon, pancreatic glucagon and GLP-1 content were measured using glucagon ELISA (EZGLU-30K, Merck Millipore), or RIA kit (250-tubes GL-32K, Millipore, USA) and GLP-1 ELISA kit Active (EGLP-35K, Millipore, MA, USA), respectively.

Sarnobat D., Moffett C.R., Tanday N., Reimann F., Gribble F.M., Flatt P.R, & Tarasov A.I. (2020). Antidiabetic drug therapy alleviates type 1 diabetes in mice by promoting pancreatic α-cell transdifferentiation. Biochemical pharmacology, 182, 114216.

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Multimodal Metabolic Assessment in Rats

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Blood glucose was measured using glucometer, unless reading exceed glucometer capabilities, in which case, plasma glucose was measured using a glucose analyzer (Analox GM9, Stourbridge, UK). Fed and fasted plasma insulin was measured via ELISA following manufacturer’s protocol (Cat. # 80-INSRT-E01, Alpco, Salem, NH, USA). Circulating leptin at 8 week, 16 week, and 24 week timepoints was measured using multiplex assay following manufacturer’s protocol (Cat. # RECYTMAG-65K, Millipore, Burlington, MA, USA). Vena cava leptin and portal glucagon-like peptide-1 (GLP-1) from control and OFS-treated rats was determined by ELISA following manufacturer’s protocol (Cat. #EGLP-35K, Millipore; Cat. # EZRL-83K, Millipore).

Weninger S.N., Ding A., Browne E.N., Frost M.L., Schiro G., Laubitz D, & Duca F.A. (2023). Longitudinal Characterization of the Gut Microbiota in the Diabetic ZDSD Rat Model and Therapeutic Potential of Oligofructose. Metabolites, 13(5), 660.

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Amish Metabolic Biomarker Protocol

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Screening blood samples were obtained by a research nurse during home visits and collected in test tubes as appropriate for each assay. Blood samples were transported to the clinical laboratory at the Amish Research Clinic (maximum transport time, 2 h) at room temp or on ice, as appropriate. After centrifugation (3300 rpm for 10 min), plasma/serum was sent on the same day to Quest Diagnostics for assay. Blood samples for OGTTs were collected in EDTA‐containing tubes for the measurement of plasma insulin and in EDTA/oxalate‐containing tubes for the measurement of plasma glucose. Fifty microliters of aprotinin solution (20,000 KIU/mL) was added per mL blood for glucagon assays. A DPP4 inhibitor (Millipore) was added to plasma samples for assays of GLP1 and GIP at a concentration of 10 μL DPP4 inhibitor per mL blood. Glucose was measured in duplicate using a YSI glucose analyzer. Insulin was assayed in duplicate using ELISA following the manufacturer's instructions using reagents in kits purchased from Mercodia (Mercodia #10‐1113‐01), assay range 3–200 mU/L. The following ELISA kits were used for assays of other peptides: intact GLP1 (Millipore #EGLP‐35K), range 2–100 pM, total GIP (Millipore #EZHGIP‐54K), range 4.2–2000 pg/mL, and glucagon (Mercodia #10‐1271‐01), assay range 1.5–130 pmol/L.

Beitelshees A.L., Streeten E.A., Shahidzadeh Yazdi Z., Whitlatch H.B., Mitchell B.D., Shuldiner A.R., Montasser M.E, & Taylor S.I. (2024). Acute pharmacodynamic responses to sitagliptin: Drug‐induced increase in early insulin secretion in oral glucose tolerance test. Clinical and Translational Science, 17(5), e13809.

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Measuring GLP-1 Secretion in NCI-H716 Cells

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NCI-H716 cells were grown in medium with or without 10 uM Olaparib. After 48 hours, supernatants were replaced by PBS containing 100 uM dipeptidyl peptidase IV inhibitor, diprotin A. Cells were subsequently stimulated for 30 minutes with or without 16 mM glucose. GLP-1 was measured by ELISA (EGLP-35K; Millipore, Billerica, MA) according to the manufacturer’s instructions.
Figure 1.High glucose conditions shorten <i>C. elegans</i> lifespan

Wild-type N2 C. elegans cultured under standard and high glucose conditions and subsequent lifespan assays. (A) Determination of lifespan for N2 worms unexposed or exposed to 16 mM glucose (P<0.002, log-rank test, n=100); (B) Determination of lifespan for N2 worms unexposed or exposed to 25 mM glucose (P<0.0001, log-rank test, n=100).

Xia Q., Lu S., Ostrovsky J., McCormack S.E., Falk M.J, & Grant S.F. (2018). PARP-1 Inhibition Rescues Short Lifespan in Hyperglycemic C. Elegans And Improves GLP-1 Secretion in Human Cells. Aging and Disease, 9(1), 17-30.

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Measuring Serum Glp-1 Levels via ELISA

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The serum level of glucagon-like peptide (Glp-1) was measured by an ELISA kit (Millipore EGLP-35k) following the manufacturer’s protocol. Before the collection of blood samples, 10 μl of DPP-IV inhibitor was added to the tubes to inhibit the degradation of Glp-1.

Hou Y.F., Shan C., Zhuang S.Y., Zhuang Q.Q., Ghosh A., Zhu K.C., Kong X.K., Wang S.M., Gong Y.L., Yang Y.Y., Tao B., Sun L.H., Zhao H.Y., Guo X.Z., Wang W.Q., Ning G., Gu Y.Y., Li S.T, & Liu J.M. (2021). Gut microbiota-derived propionate mediates the neuroprotective effect of osteocalcin in a mouse model of Parkinson’s disease. Microbiome, 9, 34.

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