Ezrgra 90k

Manufactured by Merck Group

Sourced in United States

The EZRGRA-90K is a general-purpose laboratory centrifuge with a maximum speed of 9,000 RPM and a maximum capacity of 90 milliliters. It is designed for a variety of common laboratory applications that require separation of materials based on density differences.

Automatically generated - may contain errors

Lab products found in correlation

Ezrgrt 91k, by Merck Group (4 mentions) Ezrmgh 45k, by Merck Group (3 mentions) Ezrmi 13k, by Merck Group (2 mentions) Ezml 82k, by Merck Group (2 mentions) Pefabloc, by Merck Group (2 mentions) P hydroxymercuribenzoic acid, by Merck Group (2 mentions) Victor x3, by PerkinElmer (1 mentions) Ultrasensitive mouse insulin elisa, by Mercodia (1 mentions) Glucagon elisa, by Mercodia (1 mentions) Pefabloc sc, by Merck Group (1 mentions) Hydrochloric acid (hcl), by Avantor (1 mentions) Ab108821, by Abcam (1 mentions) Akt and p akt, by Cell Signaling Technology (1 mentions) Ghsr1a, by Santa Cruz Biotechnology (1 mentions) Gapdh, by Proteintech (1 mentions) Tissuelyser 2, by Qiagen (1 mentions) Complete mini protease inhibitor cocktail, by Roche (1 mentions) Powerwave xs microplate spectrophotometer, by Agilent Technologies (1 mentions) Kc4 junior software, by Agilent Technologies (1 mentions) 22 ig1ms e01, by ALPCO (1 mentions)

13 protocols using ezrgra 90k

Plasma Ghrelin Quantification Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

Blood samples were collected from the tail vein using EDTA-treated capillaries and immediately transferred to ice-cold tube containing serine proteinase inhibitor (Pefabloc SC, 1 mg mL⁻¹ blood) and centrifuged. Then, plasma was acidified with HCl (0.05 M, final concentration) and stored in −80°C until assay. Plasma total- and acyl-ghrelin levels were determined using specific ELISA kits (EZRGRT-91K and EZRGRA-90K, respectively, Millipore, Billerica, MA).

Karasawa H., Pietra C., Giuliano C., Garcia-Rubio S., Xu X., Yakabi S., Taché Y, & Wang L. (2014). New ghrelin agonist, HM01 alleviates constipation and L-dopa-delayed gastric emptying in 6-hydroxydopamine rat model of Parkinson’s disease. Neurogastroenterology and motility : the official journal of the European Gastrointestinal Motility Society, 26(12), 1771-1782.

+ Open protocol

+ Expand

Serum Metabolic Biomarkers in Mice

Check if the same lab product or an alternative is used in the 5 most similar protocols

At the end of the 7th week of treatment, blood samples of approximately 0.6~0.9 mL were collected from abdominal artery of ether anesthetized mice after 16 hours of fasting. The blood sample was centrifuged at 2,000 ×g for 10 min at 4°C. After centrifugation, serum of each group was pooled, divided, and stored at −70°C until being analyzed. Enzyme-linked immunosorbent assay (ELISA) kits were used to determine the levels of leptin (#EZML-82K, Millipore Co., USA), active ghrelin (#EZRGRA-90K, Millipore Co., USA), and insulin (#EZRMI-13K, Millipore Co., USA). Measurements were performed using microplate leader (VictorTM X3, PerkinElmer Inc., USA) with software version of PerkinElmer 2030.

Yimam M., Jiao P., Hong M., Brownell L., Lee Y.C., Hyun E.J., Kim H.J., Kim T.W., Nam J.B., Kim M.R, & Jia Q. (2016). Appetite Suppression and Antiobesity Effect of a Botanical Composition Composed of Morus alba, Yerba mate, and Magnolia officinalis. Journal of Obesity, 2016, 4670818.

+ Open protocol

+ Expand

Fasting and Refeeding Plasma Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

Plasma samples were obtained through a submandibular puncture under isoflurane anesthesia at the end of 24 h of fasting and 6 h of refeeding. For acyl ghrelin and total ghrelin essays, plasma was treated with 1 mg/mL pefabloc SC (Sigma-Aldrich, St. Louis, MO, USA) and acidified with 0.05 N HCl. ELISA analyses were performed using the following assays: EZRGRA-90K and EZRGRT-91 K for acyl and total ghrelin (Millipore, Billerica, MA, USA), Ultrasensitive Mouse insulin ELISA (10-1247-01; Mercodia, Winston-Salem, USA), glucagon ELISA (10-1281-01; Mercodia), leptin ELISA (EZML-82K; Millipore) and GLP-1 ELISA (81508; CrystalChem).

Barahona M.J., Langlet F., Labouèbe G., Croizier S., Picard A., Thorens B, & García-Robles M.A. (2022). GLUT2 expression by glial fibrillary acidic protein-positive tanycytes is required for promoting feeding-response to fasting. Scientific Reports, 12, 17717.

+ Open protocol

+ Expand

Plasma Ghrelin Measurement and Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

Whole blood was collected into chilled, pre-coated K₃ EDTA tubes (MiniCollect, Grenier Bio-One) and immediately treated with Pefabloc (Sigma) to a final concentration of 1 mM. Blood was centrifuged at 1600 rcf for 15 minutes. Plasma was aliquoted and stored at −80°C for later use. Plasma active (acylated) ghrelin was measured by mouse/rat ELISA (EZRGRA-90K, Millipore). Plasma total ghrelin was measured by mouse/rat ELISA (EZRGRT-91K, Millipore). All samples were run in duplicate. The ratio of acylated (active form of ghrelin)/ total ghrelin was calculated by dividing the mean of the acylated ghrelin concentration by the mean of the total ghrelin concentration per state. The SEM of the ratio was calculated by simple error propagation given by the formula:

σ_{ratio} = sqrt ((σ A / A) 2 + (σ B / B) 2)) * A / B

where A and B are mean values of active (acylated) and total ghrelin, respectively.

Mohr S.M., Pra R.D., Platt M.P., Feketa V.V., Shanabrough M., Varela L., Kristant A., Cao H., Merriman D.K., Horvath T.L., Bagriantsev S.N, & Gracheva E.O. (2024). Hypothalamic hormone deficiency enables physiological anorexia. bioRxiv.

+ Open protocol

+ Expand

Ghrelin and Corticosterone in HFD Mice

Check if the same lab product or an alternative is used in the 5 most similar protocols

In a subset of animals (mice from experiment 3, exposed for 8 weeks to a standard diet or HFD, three days following MK0677 administration), blood was collected from the heart following respiratory arrest. Pefabloc (1 mg/ml, Sigma-Aldrich) was added immediately following blood collection. Plasma was subsequently obtained through centrifugation at 2500 g for 15 min at 4 °C.
Samples for ghrelin analysis were acidified with hydrochloric acid (final concentration of 0.05 N, VWR, International, Radnor, Pennsylvania, US).
Enzyme-linked immunosorbent assays were performed for acylated ghrelin (EZRGRA-90K, Millipore, Missouri, USA), total ghrelin (EZRGRT-91K, Millipore) and corticosterone (ab108821, Abcam, Cambridge, UK) concentrations according to the manufacturer's instructions.

Pierre A., Regin Y., Van Schuerbeek A., Fritz E.M., Muylle K., Beckers T., Smolders I.J., Singewald N, & De Bundel D. (2019). Effects of disrupted ghrelin receptor function on fear processing, anxiety and saccharin preference in mice. Psychoneuroendocrinology, 110.

+ Open protocol

+ Expand

Acylated Ghrelin Quantification in Mice

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Due to limitations of mouse body size and total blood volume, only the final, post-stress blood sample was used for analysis of acyl (active) ghrelin. Immediately after collection, plasma samples (50 μl) were treated with HCl to achieve a final sample concentration of 0.5 N and with 1 mg/ml protease inhibitor cocktail (AEBSP), as recommended by the kit manufacturer and similar to published methods (Meyer et al., 2014 (link)). Plasma concentration of acylated ghrelin was determined using the commercially available active ghrelin enzyme immunoassay kit (EZRGRA −90K; Millipore Sigma, Burlington, MA, USA) according to the manufacturer’s instructions. The standard curve used ranged from roughly 55 to 2000 pg/ml and samples were run in duplicate. A total of 3 samples (2 non-tg and 1 tg) were below the range of detection of the assay and were assigned 55 pg/ml, which was the lowest detectable value. Ghrelin concentration is expressed in pg/ml of plasma. Plasma ghrelin levels were log-10 transformed for analysis.

Hutchcroft J.E., Geahlen R.L., Deanin G.G, & Oliver J.M. (1992). Fc epsilon RI-mediated tyrosine phosphorylation and activation of the 72-kDa protein-tyrosine kinase, PTK72, in RBL-2H3 rat tumor mast cells.. Proceedings of the National Academy of Sciences of the United States of America, 89(19), 9107-9111.

+ Open protocol

+ Expand

Acyl-Ghrelin Levels in Ethanol Exposure

Check if the same lab product or an alternative is used in the 5 most similar protocols

For analysis of circulating levels of acyl-ghrelin, blood samples were collected after the first exposure to ethanol (PD28) and the last exposure (PD43). Given that vapor exposure involved a 14-hr exposure period, blood samples were taken on PD29 and PD44. However, the PD43 time-point when exposure was initiated for both procedures will be used for data presentation purposes. To assess enduring effects of AIE on the basal levels of circulating ghrelin, blood was also drawn in adulthood at PD73. An additional adult cohort was acutely exposed to ethanol or water via intragastric gavage on PD71–72 and blood samples were drawn 1 hour following gavage. For sampling at PD28, blood (200–400 μL) was obtained from the tail vein, whereas for PD43 and PD71–73 blood draws it was obtained from the saphenous vein. Blood samples were collected in tubes containing EDTA (1.86 mg/mL) and Pefabloc (1.0 mg/mL), and then centrifuged at 10,000 RPM for 15 minutes at 4° C. Plasma was transferred into a separate tube on ice containing 10 μL 1N HCl (10.0% total volume). Samples were subsequently shipped overnight on dry ice to Washington State University for measurement of acylated ghrelin levels using commercially available ELISA assay plates for Rat/Mouse Ghrelin (EZRGRA-90K, Millipore Sigma, Darmstadt, Germany).

Healey K.L., Landin J.D., Dubester K., Kibble S., Marquardt K., Brutman J.N., Davis J.F., Swartzwelder H.S, & Chandler L.J. (2020). Effects of Ethanol on Plasma Ghrelin levels in the Rat During Early and Late Adolescence. Alcohol (Fayetteville, N.Y.), 85, 111-118.

+ Open protocol

+ Expand

Serum Ghrelin and Lactate Measurement Protocol

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

At sacrifice, 2–3 h after lights on, the serum was used for measurement of ghrelin and lactate, using a specific enzyme-linked immunosorbent assay kit^{15 (link)–20 (link)}. The low and high detection limits for ghrelin were 0.04 and 10 ng/mL (kit number EZRGRA-90K; Merck Millipore, Rosh Haayin, Israel), and the intra- and inter-assay CVs were 1.1% and 3.2%, respectively. Serum lactate was determined at the Soroka University Medical Center Biochemistry laboratory. In a subset of n = 4 of control animals, AO, and OR arterial blood gases (pH, PCO₂, PO₂, and HCO₃⁻) were determined in parallel to their serum lactate level on day 66 after surgery. Antibodies used for evaluation of the hypothalamic protein extract by western immunoblot analysis (see Supplementary Methods) were GHSR1a (Santa Cruz Biotechnology, Santa Cruz, CA, USA), Akt and p-Akt (Cell Signaling Technology, Danvers, MA, USA), soleus AMPK and p-AMPK (MP Biomedical Solon, OH, USA), and GAPDH (Proteintech, Rosemont, IL).

Segev Y., Nujedat H., Arazi E., Assadi M.H, & Tarasiuk A. (2021). Changes in energy metabolism and respiration in different tracheal narrowing in rats. Scientific Reports, 11, 19166.

+ Open protocol

+ Expand

Plasma Ghrelin Quantification Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

For plasma ghrelin levels, p-hydroxymercuribenzoic acid was added (final concentration 1 mM) to EDTA-treated blood, tubes were centrifuged, and HCl was added (final concentration 0.1 N) prior to storage at −80 °C. No further processing of the EDTA-treated blood was performed prior to GH or IGF-1 determinations. To measure pituitary GH levels, pituitaries were lysed using RIPA lysis buffer [1% Triton X-100, 150 mM NaCl, 10 mM Tris–HCl (pH 7.5), 1 mM EDTA, 1% NP-40, and complete mini Protease Inhibitor Cocktail; Roche, Mannheim, Germany] and homogenized using a Qiagen TissueLyser II (Germantown). Ghrelin, GH, and IGF-1 levels were measured using commercial ELISA kits (EZRGRA-90K, EMD Millipore, Billerica, MA, USA; EZRMGH-45K, EMD Millipore; 22-IG1MS-E01, ALPCO, Reutlingen, Germany). The endpoint calorimetric assays were conducted using a PowerWave XS Microplate spectrophotometer and KC4 Junior software (BioTek Instruments, Winooski, VT, USA) or ClarioStar software version 5.40R2 and Mars software version 3.3 (BMG LabTech, Ortenberg, Germany).

Torz L.J., Osborne-Lawrence S., Rodriguez J., He Z., Cornejo M.P., Mustafá E.R., Jin C., Petersen N., Hedegaard M.A., Nybo M., Damonte V.M., Metzger N.P., Mani B.K., Williams K.W., Raingo J., Perello M., Holst B, & Zigman J.M. (2020). Metabolic insights from a GHSR-A203E mutant mouse model. Molecular Metabolism, 39, 101004.

+ Open protocol

+ Expand

Serum Acyl-Ghrelin Measurement Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

For serum acyl-ghrelin levels, 4-(2-aminoethyl)benzenesulfonyl fluoride hydrochloride (AEBSF) was added (final concentration 1 mg/mL) to the collected blood. We did not acidify the sample because it has been shown that HCl addition to AEBSF-treated samples does not provide enhanced hormone stability [33 (link)]. No further processing of either the EDTA-treated or the serum intended blood was performed prior to leptin, GH, IGF-1, or insulin determinations. In general, tubes were centrifuged to obtain either plasma or serum, which were then aliquoted and stored at −80 °C until processing. Leptin (#EZML-82K; Merck Millipore, Darmstadt, Germany), GH (#EZRMGH-45K; Merck Millipore), IGF-1 (#80574; Crystal Chem, Zaandam, The Netherlands), active ghrelin (#EZRGRA-90K; Merck Millipore), and insulin (#EZRMI-13K; Merck Millipore) levels were measured using commercial ELISA kits following the manufacturer's instructions. Samples were thawed only once.

Peris-Sampedro F., Stoltenborg I., Le May M.V., Zigman J.M., Adan R.A, & Dickson S.L. (2021). Genetic deletion of the ghrelin receptor (GHSR) impairs growth and blunts endocrine response to fasting in Ghsr-IRES-Cre mice. Molecular Metabolism, 51, 101223.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!