Bead ruptor 24

Manufactured by Biotage

The Bead Ruptor 24 is a high-performance homogenizer designed for efficient cell disruption. It utilizes high-speed bead agitation to effectively lyse a wide range of sample types, including bacteria, fungi, and animal tissues. The Bead Ruptor 24 is capable of processing up to 24 samples simultaneously, making it a versatile tool for various laboratory applications.

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Lab products found in correlation

Rnalater ice frozen tissue transition solution, by Thermo Fisher Scientific (1 mentions) 1.4 mm ceramic beads, by Thermo Fisher Scientific (1 mentions) Tri reagent, by Thermo Fisher Scientific (1 mentions) Superscript 4 vilo master mix, by Thermo Fisher Scientific (1 mentions) Rlt buffer, by Qiagen (1 mentions) Faststart universal sybr green master, by Roche (1 mentions) Bradford assay, by Thermo Fisher Scientific (1 mentions) Synergy htx instrument, by Agilent Technologies (1 mentions) Biotinylated anti ige detection antibody, by BD (1 mentions)

2 protocols using bead ruptor 24

RNA Extraction and qRT-PCR from Mouse Tissues

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For RNA extraction from mouse tissue, freshly collected tissue samples were flash-frozen and transferred to RNAlater-ICE Frozen Tissue Transition Solution (Invitrogen). After soaking overnight at −20°C, the tissue samples were homogenized in vials containing 1.4 mm ceramic beads (Fisherbrand) and 400 µl RLT buffer (Qiagen) using a bead mill (Bead Ruptor 24, Biotage). 200 µl of the tissue homogenate was mixed with 1 ml of TRI Reagent (Invitrogen). For extraction of RNA from cultured cells, the cell pellet was directly resuspended in TRI Reagent. Total RNA extraction was performed according to the manufacturer’s protocol. The resulting RNA was treated with 2U DNaseI enzyme (NEB) for 30 min at 37°C, followed by acidic phenol extraction and isopropanol precipitation. To generate cDNA, about 200 ng of RNA was used in a reverse transcription reaction with SuperScript IV VILO Master Mix (Invitrogen). To measure the relative expression levels of mRNAs by RT-qPCR, FastStart Universal SYBR Green Master (ROX) from Roche was used together with gene-specific primers listed in Supplementary file 1. GAPDH/Gapdh was used as housekeeping gene.

Mitschka S, & Mayr C. (2021). Endogenous p53 expression in human and mouse is not regulated by its 3′UTR. eLife, 10, e65700.

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Quantifying Immune Markers in Intestinal Tissue

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Fecal calproctectin levels were determined by ELISA using the S100A9/Calprotectin Mouse kit. Briefly, stool was homogeneized in extraction buffer [0.1 M Tris, 0.15 M NaCl, 1.0 M urea, 10 mM CaCl₂, 0.1 M citric acid monohydrate, 5 g/L BSA, pH 8.0] at 100 mg/mL and vortexed at maximum speed for 10 minutes. Diluted supernatants were used for ELISA according to manufacturer’s protocol. For multiplex cytokine assay, whole intestinal tissue was snap-frozen in liquid N₂. Tissue was then thawed into 200µl Cell extraction buffer supplemented with cOmplete mini protease inhibitor tablets and 1mM PMSF. Tissues were homogenized in Soft Tissue homogenizing tubes with a Bead Ruptor 24 (Biotage) [Output: 6, On time: 30 sec, Off time: 5 sec, Cycles: 4]. After centrifugation to remove particulate debris, total protein was quantified using a Bradford assay (ThermoFisher, 23200). A custom ProcartaPlex 18-plex assay was carried out according to manufacturer instructions and read on a Luminex 200 (Luminex Corp) instrument. Standard curves were generated and values interpolated using R. Serum IgA concentrations were measured using SBA Clonotyping System-HRP per manufacturer instructions. IgE was quantified using biotinylated anti-IgE detection antibody (BD Pharmingen,553414) and streptavidin-conjugated HRP. ELISAs were read at OD 450 on a Synergy HTX instrument (BioTek).

Campbell C., Dikiy S., Bhattarai S.K., Chinen T., Matheis F., Calafiore M., Hoyos B., Hanash A., Mucida D., Bucci V, & Rudensky A.Y. (2018). Extrathymically generated regulatory T cells establish a niche for intestinal border-dwelling bacteria and affect physiologic metabolite levels. Immunity, 48(6), 1245-1257.e9.

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