Gentlemacs m tube

Manufactured by Miltenyi Biotec

Sourced in Germany, United States, France

The GentleMACS M tubes are part of the GentleMACS Dissociator system, designed for the gentle dissociation and homogenization of tissue samples. The tubes are made of durable materials and feature a unique geometry to ensure efficient processing of tissue samples.

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Lab products found in correlation

Gentlemacs dissociator, by Miltenyi Biotec (28 mentions) Gentlemacs octo dissociator, by Miltenyi Biotec (10 mentions) Rneasy mini kit, by Qiagen (9 mentions) Rneasy plus mini kit, by Qiagen (4 mentions) Panta antibiotic mixture, by BD (3 mentions) 7h11 plates, by BD (3 mentions) High capacity cdna reverse transcription kit, by Thermo Fisher Scientific (3 mentions) Rlt buffer, by Qiagen (3 mentions) T per tissue protein extraction reagent, by Thermo Fisher Scientific (3 mentions) Glycerol, by Merck Group (2 mentions) Nanodrop 2000, by Thermo Fisher Scientific (2 mentions) Protease inhibitor cocktail, by Merck Group (2 mentions) Rneasy kit, by Qiagen (2 mentions) Trizol, by Thermo Fisher Scientific (2 mentions) Pcr fluorescence probing, by Daan Gene (2 mentions) Trizol reagent, by Thermo Fisher Scientific (2 mentions) Difco middlebrook 7h9, by BD (2 mentions) Bbl middlebrook adc enrichment, by BD (2 mentions) Gentlemacs, by Miltenyi Biotec (2 mentions) Rnalater solution, by Thermo Fisher Scientific (2 mentions)

Close spelling variants of this product

GentleMACS (287 citations) M tubes (64 citations) GentleMACS tubes (29 citations) GentleMACS dissociator and M-tubes (5 citations) GentleMACS dissociator M tubes (3 citations)

81 protocols using gentlemacs m tube

Transcriptomic Analysis of C. perfringens Infection

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Mice were intramuscularly injected with 1 × 10⁷ CFU of C. perfringens, and C. perfringens-infected femoral muscles were isolated 1.5 h after the infection. The isolated muscles were cut into small pieces of 2–4 mm in lysis buffer RLT of an RNeasy mini kit (QIAGEN, Hilden, Germany) and dissociated in a gentleMACS M tube (Miltenyi Biotec, Bergisch Gladbach, Germany) using a gentleMACS dissociator. Total RNA was extracted using the RNeasy mini kit, and the quality of purified RNA was assessed by Filgen Incorporated (Aichi, Japan). Microarray analysis was also performed by Filgen Incorporated using the Clariom S Assay for mice (Thermo Fisher Scientific Incorporated, MA, USA) and GeneChip Scanner 3000 7G (Thermo Fisher Scientific Incorporated, MA, USA). Scan data were analyzed using a software package (Expression Console Software; Thermo Fisher Scientific Incorporated, MA, USA).

Takehara M., Kobayashi K, & Nagahama M. (2021). Toll-Like Receptor 4 Protects Against Clostridium perfringens Infection in Mice. Frontiers in Cellular and Infection Microbiology, 11, 633440.

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Bee Colony Homogenization Protocol

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Pooled samples of 30 bees representing a single colony (n = 24) were transferred to a 50 ml sterile gentleMACS™ M Tube (Miltenyi Biotec Inc. Auburn, CA, USA), 15 ml of sterile 1X PBS was added and the samples were dissociated using the gentleMACS Dissociator (V1.02, Miltenyi Biotec Inc. Auburn, CA, USA, RNA_02.01). Following centrifugation at 4,700x g, for 5 minutes, at room temperature (RT), 2 ml of the homogenate were transferred to a sterile 2 ml centrifuge tube, and centrifuged at 21,100x g, for 5 minutes, at RT. A 200 μl aliquot of the homogenate was transferred to a sterile 1.5 ml Eppendorf tube for subsequent analysis. All samples were stored at -80°C prior to proceeding to the next step.

Nikulin S.L., Hesketh-Best P.J., Mckeown D.A., Spivak M, & Schroeder D.C. (2024). A semi-automated and high-throughput approach for the detection of honey bee viruses in bee samples. PLOS ONE, 19(3), e0297623.

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Quantification of Cytokines and Chemokines

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Cytokine and chemokine concentrations (pg/ml) were quantified in footpad and serum samples. For footpad samples, animals were anesthetized with ketamine-xylazine and intracardially perfused with PBS. The right paw was cut at the ankle and placed in a gentleMACS M tube (Miltenyi) filled with 1.5 ml of RIPA buffer (50 mM Tris–HCl, pH 7.4, 1% NP-40, 0.25% sodium deoxycholate, 150 mM NaCl, and 1 mM EDTA) complemented with 1× cOmplete Protease Inhibitor Cocktail (Roche). Samples were lysed in a Xiril Dispomix Tissue Homogenizer, centrifuged and the supernatants transferred into clean 2-ml microcentrifuge tubes for sonication in a Branson Ultrasonics Sonifier S-450 (70% intensity × 15 s). For serum samples, blood was collected from the retro-orbicular sinus using a glass Pasteur pipette and allowed to clot for 30 min at room temperature. Clotted blood was centrifuged at 14,000 rpm for serum isolation. Footpad lysates and serum samples were analyzed using the Cytokine & Chemokine 36-Plex Mouse ProcartaPlex Panel 1A (Thermo Fisher Scientific) according to the manufacturer’s protocol. Human plasma samples were analyzed using the Cytokine/Chemokine/Growth Factor 45-plex Human ProcartaPlex Panel 1 (Thermo Fisher Scientific). Data were acquired with Luminex FLEXMAP 3D instrument (Millipore) using xPONENT 4.0 software and analyzed with Bio-Plex Manager 6.1.1 (Bio-Rad Laboratories).

Torres-Ruesta A., Teo T.H., Chan Y.H., Amrun S.N., Yeo N.K., Lee C.Y., Nguee S.Y., Tay M.Z., Nosten F., Fong S.W., Lum F.M., Carissimo G., Renia L, & Ng L.F. (2022). Malaria abrogates O’nyong–nyong virus pathologies by restricting virus infection in nonimmune cells. Life Science Alliance, 5(4), e202101272.

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Quantification of Plasma MIF Levels

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Plasma was separated from blood samples by centrifugation at 5,000g at 4°C for 5min and stored at −80°C until use. Cecal tissue samples were rinsed gently in PBS to remove cecal contents and frozen immediately by immersing in liquid nitrogen. Lysates were prepared in gentleMACS^™ M tube (Miltenyi Biotec, CA, USA) containing lysis buffer. A manufacturer defined program on gentleMACS tissue dissociator for homogenization of intestine was used to prepare the lysates. Homogenate was centrifuged at 13,000g for 5min and supernatant was collected and stored at −80°C for further use. Levels of MIF were measured by ELISA using a protocol provided by the manufacturer (R&D Systems, MN, USA). Concentration of MIF in patient plasma samples was measured on a Bio-Plex 200 suspension array system using BioPlex Pro Human MIF magnetic beads (Bio-Rad, Hercules, CA, USA).

Jose S., Mukherjee A., Abhyankar M.M., Leng L., Bucala R., Sharma D, & Madan R. (2018). Neutralization of macrophage migration inhibitory factor improves host survival after Clostridium difficile infection. Anaerobe, 53, 56-63.

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Tissue Homogenization and Protein Extraction

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Forty grams of either whole lung tissue or tumor were placed in a gentleMACS™ M tube (Miltenyi Biotec, Auburn, CA), with 400 μL of 1x cell lysis buffer (Cell Signaling, Danvers, MA), 1 mM PMSF and protease inhibitors (Halt protease inhibitor cocktail kit, Pierce, Rockford, IL, USA), homogenized with a gentleMACS™ dissociator (Miltenyi Biotec, Auburn, CA) and then centrifuged for 5 min at 4,000 × g at 4°C. The supernatants were collected and protein concentration determined by a modified Bradford method (Bio-Rad Laboratories, Inc., Hercules, CA). Lysates were stored at −80°C.

Mariotti E.T., Premanandan C, & Lorch G. (2014). Canine pulmonary adenocarcinoma tyrosine kinase receptor expression and phosphorylation. BMC Veterinary Research, 10, 19.

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Quantifying mAb Lung Deposition

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To determine if our mAb was reaching the lungs of the animals we administered PhtD3 mAb as described above. Lungs were collected 24 hrs after mAb administration and rinsed in PBS to remove excess blood and then homogenized using gentleMACS M tube (Miltenyi Biotec). Homogenates were spun down at 4000xg for 10 minutes. Supernatants and serum were collected and serially diluted. 384-well plates were treated with 2 μg/ml of antigen in PBS for 1 h at 37 °C or overnight at 4 °C. Following this, plates were washed once with distilled water before blocking for 1 hr with 2% nonfat milk–2% goat serum in 0.05% PBS-Tween (PBS-T) (blocking buffer). Plates were washed with water three times before serially diluted samples in PBS were applied for 1 hr. Following this, plates were washed with water three times before application of 25 μL of secondary antibody (goat anti-human IgG Fc; Meridian Life Science) at a dilution of 1:4,000 in blocking solution. After incubation for 1 h, the plates were washed five times with PBS-T, and 25 μL of a PNPP (p-nitrophenyl phosphate) solution (1 mg/ml PNPP in 1 M Tris base) was added to each well. The plates were incubated at room temperature for 1 hr before reading the optical density at 405 nm on a BioTek plate reader.

Gingerich A.D., Royer F., McCormick A.L., Scasny A., Vidal J.E, & Mousa J.J. (2022). Synergistic protection against secondary pneumococcal infection by human monoclonal antibodies targeting distinct epitopes. Journal of immunology (Baltimore, Md. : 1950), 210(1), 50-60.

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Preparation of BCG Connaught Strain

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Mycobacterium bovis BCG Connaught strain was grown in 7H9 Middlebrook culture medium, supplemented with 0.4% glycerol (Sigma, USA) and 10% ADC enrichment (Becton Dickinson, France). After 2–3 weeks of growth, an aliquot of the culture was harvested and washed in 1× Dulbecco’s phosphate-buffered saline (DPBS; Gibco, France) for 8 min at 4,000 rpm. The bacterial pellet was resuspended in 5 ml of DPBS and then transferred to a GentleMACS M tube (Miltenyi Biotec, France). The RNA_01.01 protocol of the GentleMACS Dissociator (Miltenyi Biotec) was used to disrupt aggregated bacteria. The optical density of the bacteria was read at 600 nm (1 OD = 1 × 10⁸ CFU/ml), and bacteria was adjusted to the predetermined concentration and immediately used.

Bisiaux A., Boussier J., Duffy D., Quintana-Murci L., Fontes M, & Albert M.L. (2017). Deconvolution of the Response to Bacillus Calmette–Guérin Reveals NF-κB-Induced Cytokines As Autocrine Mediators of Innate Immunity. Frontiers in Immunology, 8, 796.

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RNA Extraction from MLNB and MLN FNA Samples

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RNA extraction from both MLNB and MLN FNA samples was performed using the RNAqueous 4-PCR extraction kit (Ambion). All materials form part of the RNAqueous 4-PCR kit unless otherwise stated. All equipment was wiped with RNAse ZAP (Ambion). Tubes and pipette tips (Sarstedt) were RNAse and DNAse free. MLNB samples were cut into portions not larger than 0.075 g, added to a gentleMACS M tube (Miltenyi Biotech) with 700 µl lysis buffer and homogenised by a gentleMACS Dissociator (Miltenyi). The M tube was centrifuged at 100 g at 4ºC for 10 mins, to (Continued) (Continued) ensure all material was lysed, and this step was repeated if required. RNAse-free and DNAse-free phosphate buffered saline solution (Sigma) at 4ºC was added to the MLN FNA sample in saline and centrifuged at 100 g at 4ºC. The resulting supernatant was discarded and the pellet resuspended in 250 µl lysis buffer. Lysed MLNB and MLN FNA samples were kept on ice at all times. The extractions were performed as per the manufacturer's instructions, followed by DNAse treatments to remove contaminating DNA. The RNA extract was transferred to a 1.5 ml tube (Sarstedt) and stored at -80ºC. The RNA extract underwent quantification and quality analysis using a Nanodrop 2000 (Thermo Fisher Scientific).

Dunbar D., Kwok W., Graham E., Armitage A., Irvine R., Johnston P., McDonald M., Montgomery D., Nicolson L., Robertson E., Weir W, & Addie D.D. (2019). Diagnosis of non-effusive feline infectious peritonitis by reverse transcriptase quantitative PCR from mesenteric lymph node fine-needle aspirates. Journal of feline medicine and surgery, 21(10).

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Tissue Harvest and Luciferase Assay

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Animals were euthanized and liver, heart, lung and tibialis anterior muscle were collected into gentleMACS M Tubes (Miltenyi Biotec, 130-093-236) in 3–6 ml of DMEM, and lysates were made using a gentleMACS Octo Dissociator (Miltenyi Biotec, 130-095-937). Tissue weights were calculated to normalize luciferase activity by tissue weight. Luciferase activity was assessed using the Luciferase Assay System (Promega, E1500).

Zengel J., Wang Y.X., Seo J.W., Ning K., Hamilton J.N., Wu B., Raie M., Holbrook C., Su S., Clements D.R., Pillay S., Puschnik A.S., Winslow M.M., Idoyaga J., Nagamine C.M., Sun Y., Mahajan V.B., Ferrara K.W., Blau H.M, & Carette J.E. (2023). Hardwiring tissue-specific AAV transduction in mice through engineered receptor expression. Nature Methods, 20(7), 1070-1081.

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Western Blot Analysis of AAVR Protein

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Organs were collected from WT, AAVR-KO and SELECTIV-WB mice after euthanasia. Organs were placed into gentleMACS M Tubes (Miltenyi Biotec, 130-093-236) with 3–5 ml of DMEM and lysates were made using gentleMACS Octo Dissociator (Miltenyi Biotec, 130-095-937). Laemmli sample buffer (Bio-Rad, 1610737) supplemented with 2-Mercaptoethanol (Bio-Rad, 1610710) was added to the lysates. Samples were headed at 95 °C for 10 min and cooled on ice before analysis by SDS–PAGE. Samples were run on 4–15% Mini-PROTEAN TGX Precast Gels (Bio-Rad, 4561086) and transferred using the Trans-Blot Turbo PVDF transfer kit (Bio-Rad, 1704272). Western blotting was performed using the primary antibody rabbit anti-AAVR at a 1:1,000 dilution (Proteintech, 21016-1-AP) and secondary antibody anti-rabbit-HRP at a 1:4,000 dilution (Genetex, GTX213110-01). GAPDH was detected on the same blots using mouse anti-GAPDH-HRP at a 1:4,000 dilution (Genetex, GTX627408-01). Blots were developed using SuperSignal West Femto Maximum Sensitivity Substrate (Thermo Scientific, 34096) or Supersignal West Dura Extended Duration Substrate (Thermo Scientific, 34076). Imaging was performed on a ChemiDoc Imaging System (Bio-Rad) and density was analyzed using Image Lab Software (Bio-Rad).

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