Mmhmag 44k

Manufactured by Merck Group

Sourced in United States

MMHMAG-44K is a laboratory equipment product from Merck Group. It is a magnetic separation device used for the isolation and purification of biomolecules, such as proteins, nucleic acids, and cells, from complex sample matrices. The core function of MMHMAG-44K is to provide a reliable and efficient method for magnetic separation and concentration of target analytes.

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Lab products found in correlation

47 adpms e01, by ALPCO (2 mentions) Adi 900 097, by Enzo Life Sciences (1 mentions) Z gel 1.1 ml serum preparation tubes, by Sarstedt (1 mentions) Halt proteinase and phosphatase inhibitor cocktail, by Thermo Fisher Scientific (1 mentions) Tissuelyser 2, by Qiagen (1 mentions) 5 mm steel bead, by Qiagen (1 mentions) T per tissue protein extraction reagent, by Thermo Fisher Scientific (1 mentions) Mje00, by R&D Systems (1 mentions) Mcx120, by R&D Systems (1 mentions) 7600 analyser, by Hitachi (1 mentions) Ezrmi 13k, by Merck Group (1 mentions) Neutral buffered formalin, by Merck Group (1 mentions) Superfrost plus glass slide, by Avantor (1 mentions) K603 100, by Abcam (1 mentions) Clarity bg1000, by Avantor (1 mentions) Mf2100, by R&D Systems (1 mentions) Tro100 1kt, by Merck Group (1 mentions) Protease inhibitor cocktail, by Merck Group (1 mentions) Mptmag 49k, by Merck Group (1 mentions) Ezrmi 13k, by Crystal Chem (1 mentions)

Spelling variants of this product

MMHMAG-44K Milliplex map mouse metabolic hormone magnetic bead panel – metabolism multiplex assay (3 citations) MILLIPLEX MAP Mouse Metabolic Hormone Magnetic Bead Panel (MMHMAG-44K; (3 citations) Milliplex mouse metabolic hormone magnetic bead panel (MMHMAG-44K, (2 citations)

16 protocols using mmhmag 44k

Insulin Resistance Evaluation in Mice

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Blood was collected from mice through retro-optical bleeding at 6 weeks post surgery after overnight fasting. Serum insulin and leptin were tested using multiplex kit (MMHMAG-44K, Millipore Corporation, 28820 Single Oak Drive, Temecula, California). Insulin tolerance was performed in each cohort at times indicated in Figure 1. The test was conducted with peritoneal injection of insulin at 0.7 U Kg⁻¹ after 4 h fasting. MOHA-IR was calculated with a formula: homeostatic model assessment-insulin resistance (mg dl⁻¹) = glucose × insulin/405.

Hao Z., Münzberg H., Rezai-Zadeh K., Keenan M., Coulon D., Lu H., Berthoud H.R, & Ye J. (2014). Leptin deficient ob/ob mice and diet-induced obese mice responded differently to Roux-en-Y bypass surgery. International journal of obesity (2005), 39(5), 798-805.

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Tissue and Serum Protein Extraction and Analysis

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Blood serum was isolated using Z-Gel 1.1 ml serum preparation tubes (41.1378.005, Sarstedt) according to the manufacturer’s instructions. Tissue for protein measurements was snap frozen in liquid nitrogen. 50–100 mg tissue were added to 10× the weight T-PER Tissue Protein Extraction Reagent (78510, ThermoFisher) containing Halt Proteinase and Phosphatase inhibitor cocktail (78440, ThermoFisher). Tissue was homogenized using a 5 mm steel bead (Qiagen) and a TissueLyser II (Qiagen) at 30 hz for 2 min (spleen, liver, adipose tissue, pancreas, thymus, kidney), 5 min (muscle, heart, lung, cecum, large intestine, small intestine) or 10 min (skin, stomach). Debris was pelleted and supernatant used for assays. Bone marrow was prepared by flushing 2 femurs with 500 ul PBS. After pelleting the cells, supernatant was used for BMEF measurements, while cells were taken up in T-PER lysis buffer, underwent one freeze-thaw cycle and debris was pelleted. For measurement of serum and tissue chemokine levels commercially available ELISA kits for CCL2/JE (MJE00, R&D) and CXCL12/SF-1 (MCX120, R&D) were used. For measurement of blood stress hormone corticosterone an ELISA kit was used (ADI-900–097, Enzo). Multiplex was performed using a kit for mouse metabolic hormones (MMHMAG-44K, Millipore) and human metabolic hormones (HMEMAG-34K, Millipore).

Jordan S., Tung N., Casanova-Acebes M., Chang C., Cantoni C., Zhang D., Wirtz T.H., Naik S., Rose S.A., Brocker C.N., Gainullina A., Hornburg D., Horng S., Maier B.B., Cravedi P., LeRoith D., Gonzalez F.J., Meissner F., Ochando J., Rahman A., Chipuk J.E., Artyomov M.N., Frenette P.S., Piccio L., Berres M.L., Gallagher E.J, & Merad M. (2019). Dietary intake regulates the circulating inflammatory monocyte pool. Cell, 178(5), 1102-1114.e17.

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Measuring Leptin Levels in Diabetic Plasma

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Blood plasma samples from all animals at 12 weeks of age (8 weeks of diabetes) were analyzed for leptin levels using the Milliplex Metabolic Magnetic Bead Panel (MMHMAG-44 K, Millipore; Billerica, MA.).

Coleman S.K., Rebalka I.A., D’Souza D.M., Deodhare N., Desjardins E.M, & Hawke T.J. (2016). Myostatin inhibition therapy for insulin-deficient type 1 diabetes. Scientific Reports, 6, 32495.

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Mouse Metabolic Biomarker Profiling

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Fasting plasma insulin levels were measured in mice fasted for 16 h using the Mouse Metabolic Hormone Magnetic Bead Panel (Metabolism Multiplex Assay, MMHMAG-44K, Millipore). Plasma TG, cholesterol and FFA concentrations were determined using enzymatic methods (Roche Diagnostics, Mannheim, Germany) with a Hitachi 7600 clinical chemistry analyser (Hitachi Ltd., Tokyo, Japan) in overnight-fasted mice. Serum AST and ALT levels were measured via ultraviolet methods (Roche Diagnostics) with a Hitachi 7600 analyser, according to the recommended protocols from the International Federation of Clinical Chemistry.

Lee M.S., Han H.J., Han S.Y., Kim I.Y., Chae S., Lee C.S., Kim S.E., Yoon S.G., Park J.W., Kim J.H., Shin S., Jeong M., Ko A., Lee H.Y., Oh K.J., Lee Y.H., Bae K.H., Koo S.H., Kim J.W., Seong J.K., Hwang D, & Song J. (2018). Loss of the E3 ubiquitin ligase MKRN1 represses diet-induced metabolic syndrome through AMPK activation. Nature Communications, 9, 3404.

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Multiplex ELISA of Glucagon and TNF-α

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The multiplex assay used was a multiple ELISA assay. Indeed, thanks to magnetic beads coated for different antibodies, it is possible to perform an ELISA for several antigens for the same sample in the same well. The multiplex assay was performed using a Milliplex kit (MMHMAG-44K, Millipore, Merck) following the manufacturer's instructions. Briefly, samples were incubated with a magnetic bead mix (directed against glucagon and TNFα) overnight at 4°C under agitation. The plate was washed three times with 1x wash buffer, and 50 μL of a solution containing detection antibodies was added to each well. The plate was incubated for 30 min under agitation at room temperature (RT). Fifty microliters of a streptavidin-phycoerythrin solution was added into each well, and the plate was incubated under agitation for 30 min at RT. The plate was washed three times, 100 μL sheath fluid was added into each well, and the plate was read using the Bio-Rad Luminex® 2000™ software. The gate settings were set according to the manufacturer's instructions (i.e., 8,000 to 15,000).

Daems C., Welsch S., Boughaleb H., Vanderroost J., Robert A., Sokal E, & Lysy P.A. (2019). Early Treatment with Empagliflozin and GABA Improves β-Cell Mass and Glucose Tolerance in Streptozotocin-Treated Mice. Journal of Diabetes Research, 2019, 2813489.

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Plasma Insulin, Leptin, and Adiponectin Levels

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Plasma insulin and leptin were measured using a Milliplex MAP kit with a mouse metabolic magnetic bead panel [catalog no. MMHMAG-44K (28 ), Millipore Corporation, Bedford, MA]. Plasma insulin was also measured by ELISA [catalog no. EZRMI-13K (29 ), Millipore Corporation]. Adiponectin was measured using a Milliplex MAP kit mouse adiponectin magnetic bead panel single plex [catalog no. MADPNMAG-70K-0 (30 ), Millipore Corporation]. Mean interassay and intra-assay coefficient of variation (CV) values for an insulin or leptin Milliplex MAP kit are 7% and 5%, respectively, and the assay standards range from 0.069 ng/mL to 50 ng/mL. Mean interassay and intra-assay CV values for insulin ELISA are 18% and 8%, respectively, and the assay standards range from 0.2 ng/mL to 10 ng/mL. Mean interassay and intra-assay CV values for an adiponectin Milliplex MAP kit are 11% and 3%, respectively, and the assay standards range from 12.2 pg/mL to 12,500 pg/mL.

Hubbard K., Shome A., Sun B., Pontré B., McGregor A, & Mountjoy K.G. (2019). Chronic High-Fat Diet Exacerbates Sexually Dimorphic Pomctm1/tm1 Mouse Obesity. Endocrinology, 160(5), 1081-1096.

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Intestinal Tract Segmentation and GLP-1 Quantification

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Sampling was carried out at the same time of day for all experiments. Blood samples were taken by cardiac puncture following euthanasia with 0.1 ml of Dipeptidyl peptidase IV (DPP-IV) inhibitor (BIO-TECHNE LTD.) per ml of blood, centrifuged at 1,000–2,000 × g for 10 min and the serum removed, aliquoted and stored at −20°C prior to analysis. The entire intestinal tract was excised, the contents removed by flushing with sterile Dulbecco’s Phosphate Buffered Saline (DPBS), prior to dividing into anatomically distinct segments (duodenum, jejunum, ileum, proximal colon and distal colon) that were fixed in 10% neutral buffered formalin (Sigma-Aldrich) for 24 h at 20–22°C followed by 24 h in 70% ethanol at 4°C. Tissues were then processed through a xylene/alcohol dehydration and clearing series followed by wax infiltration. Segments were embedded in paraffin wax prior to sectioning (5 μm) and mounting on SuperFrost^® Plus glass slides (VWR). ELISA (Millipore, MMHMAG-44K) was used to quantitate GLP-1 levels in serum.

Modasia A., Parker A., Jones E., Stentz R., Brion A., Goldson A., Defernez M., Wileman T., Ashley Blackshaw L, & Carding S.R. (2020). Regulation of Enteroendocrine Cell Networks by the Major Human Gut Symbiont Bacteroides thetaiotaomicron. Frontiers in Microbiology, 11, 575595.

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Comprehensive Metabolic Profiling Assay

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Glucose was measured using a Clarity BG1000 handheld glucometer (VWR, Radnor PA). (Insulin was measured using by ELISA following manufacturer protocol (Crystal Chem #90080). FGF21 was measured by ELISA following manufacturer protocol (R&D Systems #MF2100). Adiponectin was measured by ELISA following manufacturer protocol (ALPCO #47-ADPMS-E01). Serum triglycerides were measured using a colorimetric kit according to manufacturer protocol (MilliporeSigma TRO100–1KT). Serum cholesterol was measured using a colorimetric assay according to manufacturer protocol (BioVision #K603–100). Resistin was measured using a Luminex multiplex assay according to manufacturer protocol (EMD Millipore MMHMAG-44K). Complete blood cell counts were measured using a Hemavet blood analyzer according to manufacturer’s protocol.

MacArthur M.R., Mitchell S.J., Chadaideh K.S., Treviño-Villarreal J.H., Jung J., Kalafut K.C., Reynolds J.S., Mann C.G., Trocha K.M., Tao M., Aye Cho T.Z., Koontanatechanon A., Yeliseyev V., Bry L., Longchamp A., Ozaki C.K., Lewis C.A., Carmody R.N, & Mitchell J.R. (2022). Multiomics assessment of dietary protein titration reveals altered hepatic glucose utilization. Cell reports, 40(7), 111187.

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Measuring Leptin and Adiponectin Levels

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Leptin concentrations were determined by a multiplex assay (MMHMAG-44k, EMD Millipore, Billerica, MA, USA). The adiponectin concentration was determined by ELISA (BioVision, Inc.).

Lee J., Walter M.F., Korach K.S, & Noguchi C.T. (2020). Erythropoietin reduces fat mass in female mice lacking estrogen receptor alpha. Molecular Metabolism, 45, 101142.

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Submandibular Serum Collection and Metabolic Hormone Analysis

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Submandibular blood (100-200 µl) was collected in a Microvette CB300z blood collection tube (Kent Scientific, Torrington, CN). Serum was isolated by centrifugation for 10 min at 1800×g at 4°C using a tabletop centrifuge (PrismR; C2500-R). Serum was collected and treated with protease inhibitor cocktail (Sigma) and stored at −80°C until analysis. Serum was analyzed using a Milliplex MAP Mouse Metabolic Hormone Magnetic Bead Panel – Metabolism Multiplex assay (EMD Millipore; MMHMAG-44K).

Jacobs D.T., Silva L.M., Allard B.A., Schonfeld M.P., Chatterjee A., Talbott G.C., Beier D.R, & Tran P.V. (2016). Dysfunction of intraflagellar transport-A causes hyperphagia-induced obesity and metabolic syndrome. Disease Models & Mechanisms, 9(7), 789-798.

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