P5288

Manufactured by Merck Group

Sourced in United States

P5288 is a laboratory centrifuge used for separating and isolating components of a liquid mixture based on their density and sedimentation rate. The core function of this equipment is to provide controlled centrifugal force to enable efficient separation of substances within a sample.

Automatically generated - may contain errors

Lab products found in correlation

S0389, by Merck Group (2 mentions) G1890, by Merck Group (2 mentions) Micromanipulator, by Narishige (1 mentions) Fluorinert fc 770, by Merck Group (1 mentions) Inverted microscope, by Leica camera (1 mentions) M2 medium, by Merck Group (1 mentions) Cm3050, by Leica camera (1 mentions) Polyvinylpyrrolidone, by Merck Group (1 mentions) M3148, by Merck Group (1 mentions) Lithium chloride precipitation solution, by Thermo Fisher Scientific (1 mentions) Sp8 inverted confocal microscope, by Leica camera (1 mentions) Anti brdu agarose beads, by Santa Cruz Biotechnology (1 mentions)

6 protocols using p5288

Piezo-Assisted Intracytoplasmic Sperm Injection

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B6D2F1 females were superovulated by intraperitoneal injection of 5 i.U. Pregnant Mare Serum (PMS) and human Chorionic Gonadotropin (hCG). Oocytes were isolated from the oviducts 15 h after the last hormone injection and freed from cumulus cells by treatment with hyaluronidase. Testicular sperm were isolated from 8–13 week-old males as described [53 (link)]. Briefly, after removal of the tunica albuginea, testes were placed into 1% (m/v) PVP solution (P5288, Sigma) and cut into minute pieces. One part of the testicular suspension was throughout mixed with two parts 12% PVP solution and incubated at 16 °C until injection.
ICSI was performed at 17 °C using an inverted microscope (Leica, Wetzlar, Germany) equipped with micromanipulators (Narishige, Tokyo, Japan) and a piezo element (Eppendorf, Hamburg, Germany). Injection capillaries (PIEZO 8-15-NS, Origio, Charlottesville, VA, USA) were filled with Fluorinert (FC-770, Sigma) for proper Piezo pulse propagation. Testicular sperm were collected and injected into oocytes as described in detail by Yoshida et al. [54 (link)] with minor modifications. Here, testicular sperm were injected head to tail without prior removal of the sperm flagellum. Oocytes were cultured in M2 medium (Sigma) for injection. Surviving oocytes were cultivated in a drop-culture of KSOM (Gynemed, Lensahn, Germany) under mineral oil (Gynemed) at 37 °C and 5% CO₂.

Schneider S., Shakeri F., Trötschel C., Arévalo L., Kruse A., Buness A., Poetsch A., Steger K, & Schorle H. (2020). Protamine-2 Deficiency Initiates a Reactive Oxygen Species (ROS)-Mediated Destruction Cascade during Epididymal Sperm Maturation in Mice. Cells, 9(8), 1789.

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Cryopreservation of Tissue Samples

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Heart, skeletal muscle and liver were dissected in cold PBS and fixed overnight in 4% PFA at 4°C. Samples were then processed for cryo-sectioning by cryo-protecting them with a series of consecutive overnight washes in PBS containing increasing concentration of sucrose (Sigma, S0389) (10%, 20% and 30%) and finally embedded in embedding solution (15% sucrose, 8% gelatin (Sigma, G1890) and 1% polyvinylpyrrolidone (pvp; Sigma, P5288) in PBS). After waiting for the embedding solution to solidify, samples were frozen at −80°C. Tibias were fixed overnight in 2% PFA at 4°C and previous to its embedding were decalcified with two consecutives overnight 15% EDTA washes at 4°C. Then cryoprotected with a single wash of 20% sucrose and 1% pvp in PBS, and finally embed as described above.
Samples were sectioned 50 µm thick in the cryostate Leica CM3050 S and collected into microscopy slides. Liver sections for Oil Red-O staining were cut 7 µm thick. After sectioning, samples were let dry overnight at room temperature and stored at −20°C.

Luxán G., Stewen J., Díaz N., Kato K., Maney S.K., Aravamudhan A., Berkenfeld F., Nagelmann N., Drexler H.C., Zeuschner D., Faber C., Schillers H., Hermann S., Wiseman J., Vaquerizas J.M., Pitulescu M.E, & Adams R.H. (2019). Endothelial EphB4 maintains vascular integrity and transport function in adult heart. eLife, 8, e45863.

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Temporal Dynamics of Bark Transcriptome

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Bark was harvested 3 h, 6 h, 24 h, 72 h, 1 week and 4 weeks after treatment, from the first stem internode of four seedlings per treatment (water and MeJA). Each seedling was treated as a separate sample, therefore each treatment (e.g., MeJA treated) had four replicates for each harvest time. Immediately following harvesting the bark samples were flash frozen in liquid nitrogen and then stored at −80°C. Using a pestle and mortar, all bark samples were later ground to a powder in liquid nitrogen. For the samples collected 4 weeks posttreatment only half of the harvested bark was ground to a powder.
Total RNA was extracted from 30 to 50 mg of bark powder per sample using the MasterPure Complete DNA and RNA Purification Kit (Lucigen, MC85200). To denature ribonucleases and reduce polyphenolic contamination of extracted nucleic acids, 0.5% β‐mercaptoethanol (Sigma‐Aldrich, M3148) and 1% polyvinylpyrrolidone (PVP; Sigma‐Aldrich, P‐5288) were added to the extraction buffer. To reduce carbohydrate contamination, nucleic acids were precipitated using 0.5 volumes of 7.5 M lithium chloride precipitation solution (ThermoFisher Scientific, AM9480), subject to an incubation step at −20°C for 2 h, and centrifuged for 30 min at 16 500 × g and 4°C. Nucleic acids were resuspended in 60 μl of nuclease‐free water.

Wilkinson S.W., Dalen L.S., Skrautvol T.O., Ton J., Krokene P, & Mageroy M.H. (2022). Transcriptomic changes during the establishment of long‐term methyl jasmonate‐induced resistance in Norway spruce. Plant, Cell & Environment, 45(6), 1891-1913.

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Decalcification and Cryosection of Bone Tissues

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Freshly dissected bone tissues were fixed in ice-cold 4% paraformaldehyde solution for 4 h. After PBS washing, decalcification was carried out with 10% EDTA/Tris/HCL pH = 7.0 (15575-038; Invitrogen) at 4 °C with constant shaking for 48 h and after three times washing with PBS, decalcified bones were immersed into 20% sucrose (S0389; Sigma) and 1% polyvinylpyrrolidone (PVP) (P5288; Sigma) solution for 24 h. Finally, the tissues were embedded and frozen in 8% gelatin (porcine) (G1890; Sigma) in presence of 15% sucrose and 1% PVP. Cryosections were produced at a Leica CM3050S cryostat
Cryosection was stored at −20 °C. Before starting the staining procedure, they were allowed to acclimate to room temperature. Slides were then rehydrated in PBS and permeabilized with PBS containing 0.3% Triton X-100 by washing them three times for 10 min. Tissue was blocked for one hour in freshly prepared blocking solution (3% BSA, 0.1% Triton X-100, 20 mM MgCl₂, and 5% donkey serum in PBS). Primary antibodies were incubated overnight at 4 °C in a blocking solution. On a consecutive day, the primary antibodies were washed four times 5 min in PBS. Secondary antibodies were incubated for one hour at room temperature in PBS containing 5% BSA. Immunostainings were imaged in a Leica SP8 confocal inverted microscope.

Hoffmann J., Luxán G., Abplanalp W.T., Glaser S.F., Rasper T., Fischer A., Muhly-Reinholz M., Potente M., Assmus B., John D., Zeiher A.M, & Dimmeler S. (2021). Post-myocardial infarction heart failure dysregulates the bone vascular niche. Nature Communications, 12, 3964.

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Anti-BrdU Immunoprecipitation Bead Protocol

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Anti BrdU-agarose beads (Santa Cruz Biotechnology SC 32323, 25 μL /IP) are washed twice by adding buffer I (0.5X SSPE (0.5mM EDTA and 74.5mM NaCl in 5mM phosphate buffer (pH 7.4) with 0.05% Tween20 and 0.1% polyvinylpyrrolidone [PVP, SIGMA P5288])and spinning 2’ at 2rcf. Beads are then blocked overnight on a rotating wheel at 4°C (up to 400 μL beads in 900 Buffer I supplemented with 100 μL of 10mg/ml RNase free BSA).

Jullien J., Vodnala M., Pasque V., Oikawa M., Miyamoto K., Allen G., David S.A., Brochard V., Wang S., Bradshaw C., Koseki H., Sartorelli V., Beaujean N, & Gurdon J. (2017). Gene Resistance to Transcriptional Reprogramming following Nuclear Transfer Is Directly Mediated by Multiple Chromatin-Repressive Pathways. Molecular Cell, 65(5), 873-884.e8.

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Endophyte-Coated Bentgrass Seed Preparation

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Endophytes were cultured in LB overnight at 37 °C, shaking at 250 rpm. Cells were centrifuged, washed twice in 10 mM tris HCl (pH 7), then suspended to OD₅₉₅ = 0.5. From each suspension, 500 μl were diluted in 5 ml of 9.3 % PVP aqueous solution (P-5288, Sigma, USA) and used to coat creeping bentgrass seeds.

Shehata H.R., Lyons E.M., Jordan K.S, & Raizada M.N. (2016). Relevance of in vitro agar based screens to characterize the anti-fungal activities of bacterial endophyte communities. BMC Microbiology, 16, 8.

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