Robovials

Manufactured by Thermo Fisher Scientific

Robovials are a specialized laboratory equipment product designed for automated sample handling and storage. They provide a standardized vial system for efficient organization and management of small samples within a laboratory environment. Robovials are constructed to integrate seamlessly with various automated systems and robotic handling equipment, enabling efficient and precise sample processing and retrieval.

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Lab products found in correlation

Selexion tuning kit, by AB Sciex (4 mentions) Lipidyzer internal standard mix, by AB Sciex (3 mentions) Sciex lipidyzer platform, by AB Sciex (2 mentions) Omni bead ruptor elite, by Omni International (2 mentions) Sciex 5500, by AB Sciex (2 mentions) Lipidyzer platform, by AB Sciex (1 mentions) Speedvac spd 300dda, by Thermo Fisher Scientific (1 mentions) 2.8 mm ceramic beads, by Omni International (1 mentions) Speedvac, by Thermo Fisher Scientific (1 mentions)

7 protocols using robovials

Targeted Lipidomic Analysis of Plasma

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Lipids from 25 ul of plasma were extracted using a modified Bligh and Dyer extraction method by the UCLA lipidomics core facility⁵⁶. Prior to biphasic extraction, a 13-lipid class Lipidyzer Internal Standard Mix is added to each sample (AB Sciex, 5040156). Following two successive extractions, pooled organic layers were dried down in a Genevac EZ-2 Elite evaporator. Lipid samples were resuspended in 1:1 methanol/dichloromethane with 10mM ammonium acetate and transferred to robovials (Thermo 10800107) for analysis. Samples were analyzed on the Sciex Lipidyzer Platform for targeted quantitative measurement of 1100 lipid species across 13 classes. Differential Mobility Device on Lipidyzer was tuned with SelexION tuning kit (Sciex 5040141). Instrument settings, tuning settings, and MRM list are available upon request.

Fung T.C., Vuong H.E., Luna C.D., Pronovost G.N., Aleksandrova A.A., Riley N.G., Vavilina A., McGinn J., Rendon T., Forrest L.R, & Hsiao E.Y. (2019). Intestinal serotonin and fluoxetine exposure modulate bacterial colonization in the gut. Nature microbiology, 4(12), 2064-2073.

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Quantitative Lipid Profiling of Liver

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A total of 50–100 mg of frozen liver are homogenized in the Omni Bead Ruptor Elite with 2 mL homogenizer tube system (Omni, 19-628D). Samples are homogenized in cold phosphate-buffered saline (PBS) for 3 cycles of 10 s each at 5 m/s with a 10 s dwell time between cycles. A total of 3–6 mg of homogenized material are applied to a modified Bligh and Dyer extraction^{26 (link)}. Prior to biphasic extraction, a 13 lipid class Lipidyzer Internal Standard Mix is added to each sample (AB Sciex, 5040156). Following two successive extractions, pooled organic layers are dried down in a Genevac EZ-2 Elite. Lipid samples are resuspended in 1:1 methanol/dichloromethane with 10 mM ammonium acetate and transferred to robovials (Thermo 10800107) for analysis.
Samples are analyzed on the Sciex Lipidyzer Platform for targeted quantitative measurement of 1100 lipid species across 13 lipid sub-classes. Differential Mobility Device on Lipidyzer is tuned with SelexION tuning kit (Sciex 5040141). Instrument settings, tuning settings, and MRM list available upon request. Data analysis performed on Lipidyzer software. Quantitative values are normalized to milligrams of material used.

Zhang Z., Feng A.C., Salisbury D., Liu X., Wu X., Kim J., Lapina I., Wang D., Lee B., Fraga J., Pan C., Williams K.J., Lusis A.J., Scumpia P, & Sallam T. (2020). Collaborative interactions of heterogenous ribonucleoproteins contribute to transcriptional regulation of sterol metabolism in mice. Nature Communications, 11, 984.

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Targeted Lipidomic Analysis of Plasma

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Serum Lipidomic Profiling of NAFLD Patients

Cited 1 time

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Serum samples from 98 NAFLD cases and 100 controls with complete data were identified for lipid extraction. First, samples were thawed and 25ul of serum were pipetted into a glass tube for extraction. A modified Bligh and Dyer extraction [57 (link)] was carried out on the serum samples. Prior to biphasic extraction, the Lipidyzer Internal Standard Mix that contains 54 lipid standards across 13 subclasses, was added to each sample (AB Sciex, 5,040,156). The 13 subclasses include: cholesterol esters (CE), ceramides (CER), diacylglycerols (DAG), dihydroceramides (DCER), free fatty acids (FFA), hexosylceramides (HCER), lactosylceramides (LCER), lysophosphatidylcholines (LPC), lysophosphatidylethanolamines (LPE), phosphatidylcholines (PC), phosphatidylethanolamines (PE), sphingomyelins (SM), and triacylglycerols (TAGs). Following two successive extractions, pooling/concentrating of the organic layers were dried down in a Genevac EZ-2 Elite for direct infusion lipidomic analysis. Lipid samples were resuspended in 1:1 methanol/dichloromethane with 10 mM Ammonium Acetate and transferred to robovials (Thermo 10,800,107) for analysis. Samples were processed between November 2018 and February 2019 at the UCLA Lipidomics Laboratory.

Flores Y.N., Amoon A.T., Su B., Velazquez-Cruz R., Ramírez-Palacios P., Salmerón J., Rivera-Paredez B., Sinsheimer J.S., Lusis A.J., Huertas-Vazquez A., Saab S., Glenn B.A., May F.P., Williams K.J., Bastani R, & Bensinger S.J. (2021). Serum lipids are associated with nonalcoholic fatty liver disease: a pilot case-control study in Mexico. Lipids in Health and Disease, 20, 136.

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Lipidomic Analysis of HAEC Lysates and Conditioned Medium

Cited 2 times

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Lipidomic analysis was performed at the University of California Los Angeles (UCLA) Lipidomic Core. A modified Bligh and Dyer extraction (93 (link)) was carried out on HAEC lysates or conditioned medium aliquots. Before biphasic extraction, a standard mixture of 75 lipid standards (Avanti, 330820, 861809, 330729, 330727, and 791642) was added to each sample. Following two successive extractions, pooled organic layers were dried down in a Thermo SpeedVac SPD300DDA using ramp setting 4 at 35°C for 45 min with a total run time of 90 min. Lipid samples were resuspended in 1:1 methanol:dichloromethane with 10 mM ammonium acetate and transferred to robovials (Thermo Fisher Scientific, 10800107) for analysis.
Samples were analyzed by direct infusion on a Sciex 5500 with Differential Mobility Device (DMS) (comparable to Sciex Lipidyzer platform) with a targeted acquisition list consisting of 1450 lipid species across 17 subclasses. The DMS was tuned with EquiSPLASH LIPIDOMIX (Avanti, 330731). Data analysis was performed with in-house data analysis workflow. Instrument settings, multiple reaction monitoring (MRM) lists, and analysis method are previously described (94 (link)). Quantitative values were normalized to cell counts or medium volume.

Cavallero S., Roustaei M., Satta S., Cho J.M., Phan H., Baek K.I., Blázquez-Medela A.M., Gonzalez-Ramos S., Vu K., Park S.K., Yokota T., Sumner J., Mack J.J., Sigmund C.D., Reddy S.T., Li R, & Hsiai T.K. (2024). Exercise mitigates flow recirculation and activates metabolic transducer SCD1 to catalyze vascular protective metabolites. Science Advances, 10(7), eadj7481.

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Targeted Quantitative Lipidome Analysis

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50-100 mg of tissue were collected in tube pre-loaded with 2.8mm ceramic beads (Omni #19-628). Tissue is homogenized with PBS in the Omni Bead Ruptor Elite (3 cycles of 10 seconds at 5 m/s with a 10 second dwell time), and 3-6mg of homogenate were used for extraction (Bligh and Dyer extraction (Bligh, 1959)). Prior to biphasic extraction, a 13-lipid class Lipidyzer Internal Standard Mix is added to each sample (AB Sciex, 5040156). Following two successive extractions, pooled organic layers were dried in a Genevac EZ-2 Elite. Lipid samples were resuspended in 1:1 methanol/dichloromethane with 10mM Ammonium Acetate and transferred to robovials (Thermo 10800107) for analysis. Samples were analyzed on the Sciex Lipidyzer Platform for targeted quantitative measurement of 1100 lipid species across 13 classes. Differential Mobility Device on Lipidyzer is tuned with SelexION tuning kit (Sciex 5040141). Data analysis performed on Lipidyzer software and quantitative values were normalized to mg amounts used.

Palafox-Sánchez V., Ying Z., Royes L.F, & Gomez-Pinilla F. (2021). The Interaction between brain and liver regulates lipid metabolism in the TBI pathology. Biochimica et biophysica acta. Molecular basis of disease, 1867(4), 166078.

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Lipidomics Extraction and Profiling

Cited 2 times

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Lipid Extraction and mass spectrometry was performed by UCLA Lipidomics. A modified Bligh and Dyer extraction^{35 (link)} was done to extract lipids from samples. Prior to extraction, 70 lipid standards across 17 subclasses were added to each sample (AB Sciex 5040156, Avanti 330827, Avanti 330830, Avanti, 791642). Extraction was performed twice, and the pooled organic layers were dried by Speedvac (Thermo, SPD300DDA) with ramp setting 4, 35°C for 45 min with a total run time of 90 min. Samples were resuspended in 1:1 methanol/dichloromethane with 10 mM ammonium acetate and transferred to robovials (Thermo, 10800107) for analysis.
Samples were analyzed by direct infusion on a Sciex 5500 with differential mobility device (DMS), tuned with EquiSPLASH LIPIDOMIX (Avanti 330731) with a targeted acquisition list of 1450 lipid species. Descriptions of instruments settings, MRM list and in-house data analysis workflow is available.^{34 (link)}

Kreida S., Narita A., Johnson M.D., Tocheva E.I., Das A., Ghosal D, & Jensen G.J. (2023). Cryo-EM structure of the Agrobacterium tumefaciens type IV secretion system-associated T-pilus reveals stoichiometric protein-phospholipid assembly. Structure (London, England : 1993), 31(4), 385-394.e4.

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