Fastprep lysing matrix tubes

Manufactured by MP Biomedicals

Sourced in United States, France

The FastPrep Lysing Matrix Tubes are a product designed for efficient cell disruption and sample homogenization. These tubes contain specialized beads that help to physically break down and lyse cells, facilitating the release of cellular contents for further analysis or processing.

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Lab products found in correlation

Rneasy kit, by Qiagen (3 mentions) Fastprep 24 homogenizer, by MP Biomedicals (2 mentions) Express one step superscript qrt pcr kit, by Thermo Fisher Scientific (2 mentions) Express one step sybr greener kit, by Thermo Fisher Scientific (2 mentions) Dnase 1, by Thermo Fisher Scientific (2 mentions) Lysing matrix a, by MP Biomedicals (1 mentions) Lysing matrix d, by MP Biomedicals (1 mentions) Rlt buffer, by Qiagen (1 mentions) Supersignal west dura extended duration substrate, by Thermo Fisher Scientific (1 mentions) Anti rabbit igg hrp, by Cell Signaling Technology (1 mentions) Stella 3200, by Raytest (1 mentions) High pure template kit, by Roche (1 mentions) Magnalyser centrifuge, by Roche (1 mentions) Rneasy mini kit, by Qiagen (1 mentions) Euthanimal, by Alfasan (1 mentions) Serotonin, by Eagle Biosciences (1 mentions) Bessman tissue pulverizer, by Thermo Fisher Scientific (1 mentions) Complete mini protease inhibitor cocktail, by Roche (1 mentions)

8 protocols using fastprep lysing matrix tubes

ASO Delivery to CNS and Liver in Mice

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For central nervous system administration, a single ASO dose of 500 μg in phosphate buffered saline (PBS) was delivered by intracerebroventricular injection to 8-week-old C57BL/6 female mice (JAX). Four weeks post-treatment, whole brain and thoracic spinal cord tissue was lysed using FastPrep Lysing Matrix Tubes (MP-Biomedicals) in RLT buffer (Qiagen) containing 1% beta-mercaptoethanol and RNA was isolated using the RNeasy kit (Qiagen). No body weight change or inflammation (Aif1 qRT-PCR) was observed.
For systemic administration, 6-week-old BALB/c male mice (JAX) were dosed twice per week for 3 weeks with 50 mg/kg ASO in PBS. Liver was harvested 48 h after the last dose. Tissue was lysed using FastPrep Lysing Matrix Tubes (MP-Biomedicals) in guanidine isothiocyanate (Life Technologies) containing 8% beta-mercaptoethanol and RNA was isolated using the RNeasy kit (Qiagen). No changes in body weight, organ weight or plasma markers were observed.

Ward A.J., Norrbom M., Chun S., Bennett C.F, & Rigo F. (2014). Nonsense-mediated decay as a terminating mechanism for antisense oligonucleotides. Nucleic Acids Research, 42(9), 5871-5879.

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Oxylipin Extraction and Analysis Protocol

Cited 1 time

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Oxylipin extraction methods have been previously published ⁶⁶. Briefly, hind paw and dorsal
horn were transferred into FastPrep Lysing Matrix tubes on ice (MP Biomedicals,
USA; Lysing Matrix A for hind paw, Lysing Matrix D for dorsal horn) and ice-cold
methanol containing an antioxidant mixture were immediately added to each
sample. Internal standards were added and samples were homogenized by shaking
with a FastPrep-24 homogenizer (MP Bio). Proteins were precipitated by storing
samples at −80°C for 1 hour followed by centrifugation at 17000g
in 4°C for 10 min. Half the supernatant was transferred to a new
microfuge tube. and stored in −80 °C until solid phase extraction
(SPE) purification and liquid chromatography-tandem mass spectrometry (LC-MS/MS)
analysis of unesterified oxylipins. To analyze the total oxylipin pool, the
remaining half of the supernatant was saponified with methanolic sodium
carbonate at 60°C for 30 min under gentle shaking. The solution was then
neutralized (to pH 7) using acetic acid and stored in −80°C
overnight until SPE and LC-MS/MS analysis. SPE and LC-MS/MS procedures have been
described previously ^{66 ,105 (link)}

Domenichiello A.F., Sapio M.R., Loydpierson A.J., Maric D., Goto T., Horowitz M.S., Keyes G.S., Yuan Z.X., Majchrzak-Hong S.F., Mannes A.J., Iadarola M.J, & Ramsden C.E. (2020). Molecular pathways linking oxylipins to nociception in rats. The journal of pain, 22(3), 275-299.

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Extraction and Analysis of Lipid Metabolites

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Solid tissues (human skin, rat hind paw, or rat dorsal horn) were transferred into FastPrep Lysing Matrix tubes on ice (Lysing Matrix A for the skin and hind paw and Lysing Matrix D for the dorsal horn; MP Biomedicals), and at least eight times greater volume of ice-cold methanol with 0.02% (v/v) BHT and 0.02% (v/v) EDTA was immediately added to each tube. A known amount of internal standards was added to each sample, and samples were homogenized using a FastPrep-24 homogenizer (MP Biomedicals). Tissue homogenates were transferred to −80°C for 1 hour to precipitate proteins. Homogenates were centrifuged at 17,000g in 4°C for 10 min, and the supernatant was transferred to a new test tube. Half the supernatant was stored in −80°C until SPE purification and LC-MS/MS analysis (see above). To allow for the analysis of total lipid pools, we saponified the other half of the supernatant with 2.6% sodium carbonate (by weight) at 60°C for 30 min under gentle shaking. The solution was then neutralized (pH 5 to 7) using acetic acid and stored in −80°C overnight. Immediately before purification by SPE and LC-MS/MS analysis, lipid extracts (free and saponified total) were added to a ninefold greater volume of ice-cold water.

Ramsden C.E., Domenichiello A.F., Yuan Z.X., Sapio M.R., Keyes G.S., Mishra S.K., Gross J.R., Majchrzak-Hong S., Zamora D., Horowitz M.S., Davis J.M., Sorokin A.V., Dey A., LaPaglia D.M., Wheeler J.J., Vasko M.R., Mehta N.N., Mannes A.J, & Iadarola M.J. (2017). A systems approach for discovering linoleic acid derivatives that potentially mediate pain and itch. Science signaling, 10(493), eaal5241.

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Western Blotting of PDX Tumor Proteins

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Total protein extracts were prepared by lysing frozen PDX tumor cells in FastPrep lysing matrix tubes (MP Biomedicals, Illkirch, France) with lysis buffer containing cOmplete protease inhibitor cocktail, PhosSTOP phosphatase inhibitor (Roche, Mannheim, Germany). 30 μg protein were separated by SDS-PAGE and transferred onto nitrocellulose membranes. For immunoblotting, membranes were probed with primary antibodies overnight at 4°C, then incubated with anti-rabbit IgG-HRP (Cell Signaling). Signals were detected by enhanced chemiluminescence (SuperSignal WestDuraExtendedDurationSubstrate, Thermo Fisher Scientific) using CCD camera STELLA3200 (Raytest, Straubenhardt, Germany). Antibodies were used at a dilution of 1:1000, except anti-β-actin (loading control; 1:10000).

Mönch D., Bode-Erdmann S., Kalla J., Sträter J., Schwänen C., Falkenstern-Ge R., Klumpp S., Friedel G., Ott G, & Kalla C. (2018). A subgroup of pleural mesothelioma expresses ALK protein and may be targetable by combined rapamycin and crizotinib therapy. Oncotarget, 9(29), 20781-20794.

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Quantifying Gene Expression in Neurons

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Total cellular RNA was isolated from cultured neurons and mouse tissue using the RNeasy kit (Qiagen). For preparation of mouse tissue, samples were first lysed using FastPrep Lysing Matrix Tubes (MP-Biomedicals) in RLT buffer containing 1% β-mercaptoethanol. On-column DNase digestion was performed for all samples. For qRT-PCR, approximately 10 ng RNA was added to EXPRESS One-Step SuperScript qRT-PCR Kit (Life Technologies) with Taqman primer and probe sets or EXPRESS One-Step SYBR GreenER Kit (Life Technologies) with SYBR primer sets (see Extended Data Table 1 for sequences). All quantification was performed by the relative standard curve method and normalized to total RNA by Ribogreen or to the housekeeping genes Gapdh.

Meng L., Ward A.J., Chun S., Bennett C.F., Beaudet A.L, & Rigo F. (2014). Towards a therapy for Angelman syndrome by reduction of a long non-coding RNA. Nature, 518(7539), 409-412.

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Anolis Tissue Extraction and Analysis

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Anolis tissue samples were obtained from a healthy male Anolis carolinensis lizard that had been euthanized by intra-coelomic injection of pentobarbital (200 mg kg⁻¹ BW, Euthanimal^®, Alfasan International, The Netherlands). Organs were directly frozen in liquid nitrogen. Genomic DNA was isolated using the high pure template kit (Roche) according to the manufacturer’s instructions. RNA was extracted from tissue lysed with RLT buffer (1% β-mercaptoethanol) (Qiagen) in 1.4 mm Fastprep lysing matrix tubes (MPbio) in a Magna Lyser centrifuge (6,500 × g, 40 s, RT) (Roche). Total RNA was isolated using the RNeasy mini kit (Qiagen) following the manufacturer’s instructions, treated with DNase I (1 U mg⁻¹ RNA, Thermo Scientific) and stored at −80 °C until use.

Voogdt C.G., Bouwman L.I., Kik M.J., Wagenaar J.A, & van Putten J.P. (2016). Reptile Toll-like receptor 5 unveils adaptive evolution of bacterial flagellin recognition. Scientific Reports, 6, 19046.

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Cecal Tissue Serotonin and Glutamate Analysis

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Cecal tissue samples (100 mg) were pulverized using a Bessman Tissue pulverizer (ThermoFisher Scientific) followed by homogenization in cold lysis buffer (100-mg tissue/mL; 20-mM Tris, 0.25M sucrose, 2.0-mM EDTA, 10-mM EGTA, 1.0% Triton X-100) containing Complete Mini protease inhibitor cocktail (one tablet/10 mL; Roche Diagnostics, Indianapolis, IN) using FastPrep® Lysing Matrix tubes (MP Biomedicals). Homogenates were clarified by centrifugation at 21 000 × g for 40 minutes at 4°C. Commercially available enzyme-linked immunosorbent assay (ELISA) kits were used to determined levels of serotonin (Eagle Biosciences, Nashua, NH) and glutamate (Abnova, Taiwan) in the cecal tissue homogenate according to the manufacturer’s protocol.

Vetreno R.P., Massey V, & Crews F.T. (2019). Long-lasting microbial dysbiosis and altered enteric neurotransmitters in adult rats following adolescent binge ethanol exposure. Addiction biology, 26(1), e12869.

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Quantifying Gene Expression in Neurons

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Meng L., Ward A.J., Chun S., Bennett C.F., Beaudet A.L, & Rigo F. (2014). Towards a therapy for Angelman syndrome by reduction of a long non-coding RNA. Nature, 518(7539), 409-412.

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