Vacutainer cpt cell preparation tube

Manufactured by BD

Sourced in United States

The BD Vacutainer® CPT™ Cell Preparation Tubes are designed for the separation and collection of mononuclear cells from whole blood samples. The tubes contain a gel and density gradient medium that facilitate the isolation of mononuclear cells during centrifugation.

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Lab products found in correlation

Trizol reagent, by Thermo Fisher Scientific (3 mentions) Synthetic tipped applicator, by Thermo Fisher Scientific (3 mentions) Sodium heparin, by BD (3 mentions) Rpmi 1640, by Thermo Fisher Scientific (3 mentions) Negative selection based human b cell enrichment kit, by STEMCELL (3 mentions) Rpmi 1640 medium, by Thermo Fisher Scientific (3 mentions) Countess 1, by Thermo Fisher Scientific (2 mentions) Red blood cell lysis buffer, by BioLegend (2 mentions) Edta vacutainer blood collection tubes, by BD (2 mentions) Isogen, by Nippon Gene (2 mentions) Bioanalyzer, by Agilent Technologies (1 mentions) Rneasy mini kit, by Qiagen (1 mentions) β mercaptoethanol, by Merck Group (1 mentions) Pha m, by Thermo Fisher Scientific (1 mentions) Facscanto 2, by BD (1 mentions) Human th17 treg phenotyping kit, by BD (1 mentions) Human umbilical vein endothelial cells (huvec), by Lonza (1 mentions) Hmvec, by Lonza (1 mentions) Cell ranger v 1, by 10x Genomics (1 mentions) Chromium controller, by 10x Genomics (1 mentions)

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Vacutainer tubes (1123 citations) BD Vacutainer CPT tubes (175 citations) BD Vacutainer CPT (109 citations) BD Vacutainer Cell Preparation Tubes (67 citations) CPT tubes (67 citations) Cell Preparation Tubes (59 citations) CPT Vacutainer tubes (18 citations) BD Vacutainer Cell Preparation Tubes (CPT) (8 citations) BD Vacutainer® CPTTM (7 citations) CPT cell preparation tubes (4 citations)

75 protocols using vacutainer cpt cell preparation tube

Isolation and RNA Extraction from PBMCs

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PBMCs were prepared using BD Vacutainer CPT Cell Preparation Tubes (BD, Franklin Lakes, NJ, USA). The blood processing reported below was according to the manufacturer’s instructions. The tubes were centrifuged at room temperature for 20 min in a horizontal rotor at 1800 relative centrifugal force (RCF) within 1 h of blood collection. The plasma layer and the cells from both CPT tubes were transferred to one conical centrifuge tube. Phosphate-buffered saline was added to a final volume of 2 mL, the tubes were capped, and the cells were mixed by inversion. Subsequently, the tubes were centrifuged for 10 min at 4 °C and 1500–1800 RCF. The supernatant was aspirated and 1 mL TRIzol^® reagent (Invitrogen, Carlsbad, CA, USA) was added to isolate total RNA according to the manufacturer’s instructions. Total RNA was frozen immediately in liquid nitrogen and stored at −80 °C. The quality of total RNA was determined using microfluidic electrophoresis (Bioanalyzer; Agilent Technologies, Palo Alto, CA, USA).

Kawaguchi K., Suzuki E., Yamaguchi A., Yamamoto M., Morita S, & Toi M. (2016). Altered expression of major immune regulatory molecules in peripheral blood immune cells associated with breast cancer. Breast Cancer (Tokyo, Japan), 24(1), 111-120.

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Isolation and Lysis of Mononuclear Cells

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Blood samples from the study subjects were collected into BD Vacutainer^® CPT Cell Preparation tubes with sodium citrate (BD, Franklin Lakes, NJ). After 30 min incubation at room temperature (RT), CPT tubes were centrifuged at 1800 × g for 30 min at 22°C, and the isolated mononuclear cells were collected. They were washed (1× PBS, 2 mM EDTA), centrifuged at 800 × g for 8 min at 22°C and resuspended in red blood cell lysis buffer (155 mM NH₄Cl, 10 mM KHCO₃, 2 mM EDTA). The cells were centrifuged at 200 × g for 5 min at 22°C, and the washes with lysis buffer were repeated if necessary. The purified mononuclear cells were washed twice with washing buffer by centrifuging at 200 × g for 5 min at 22°C. The cell suspension was lysed by suspending in RLT-buffer (RNeasy^® Mini Kit; Qiagen, Hilden, Germany) containing 1%-β-mercaptoethanol (Sigma-Aldrich). The cell lysate was stored at −70°C.

Salonurmi T., Nabil H., Ronkainen J., Hyötyläinen T., Hautajärvi H., Savolainen M.J., Tolonen A., Orešič M., Känsäkoski P., Rysä J., Hakkola J, & Hukkanen J. (2020). 4β-Hydroxycholesterol Signals From the Liver to Regulate Peripheral Cholesterol Transporters. Frontiers in Pharmacology, 11, 361.

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PBMC Isolation from Venous Blood

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For each subject, 8 mL of venous blood was collected into BD Vacutainer^® CPT™ cell preparation tubes (BD Biosciences, Franklin Lakes, NJ, USA) containing 1 mL of Sodium Citrate and 2 mL of Ficoll. PBMCs were separated by gradient centrifugation according to manufacturer’s protocol. Freshly isolated PBMCs were immediately used for immunostaining or patch clamp experiments or frozen at −80 °C until RNA extraction.

Carlini V., Verduci I., Cianci F., Cannavale G., Fenoglio C., Galimberti D, & Mazzanti M. (2020). CLIC1 Protein Accumulates in Circulating Monocyte Membrane during Neurodegeneration. International Journal of Molecular Sciences, 21(4), 1484.

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Vitamin D Status Monitoring in Infant Cohort

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Plasma from blood samples was drawn into light-protected tubes (BD Vacutainer®CPT™ cell preparation tubes) and analyzed at the Institute for Health and Welfare, Helsinki, Finland. 25(OH)D concentration was measured using the ARCHITECT 25-OH vitamin D chemiluminescent microparticle immunoassay (3 (link)). The laboratory participates in the vitamin D external quality assessment scheme. Plasma samples for 25(OH)D were collected from visits at 3, 6, 9, and 12 months of age and then annually up or until and including the time of seroconversion for CDA-cases. Vitamin D status was categorized according to cut-offs previously used in pediatric populations, which are as follows: <30 nmol/L, 30–50 nmol/L, 50–75 nmol/L, and >75 nmol/L (30 (link), 31 (link)).

Andrén Aronsson C., Liu X., Norris J.M., Uusitalo U., Butterworth M.D., Koletzko S., Virtanen S.M., Erlund I., Kurppa K., Hagopian W.A., Rewers M.J., She J.X., Toppari J., Ziegler A.G., Akolkar B., Krischer J.P, & Agardh D. (2021). 25(OH)D Levels in Infancy Is Associated With Celiac Disease Autoimmunity in At-Risk Children: A Case–Control Study. Frontiers in Nutrition, 8, 720041.

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Measuring PBMC Proliferation Using CFSE

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Peripheral blood mononuclear cells (PBMC) were isolated from venous blood using BD Vacutainer CPT cell preparation tubes. Proliferation was measured using a carboxyfluorescein succinimidyl ester (CFSE) cell division assay, as previously described [25 (link)]. The stimulation index (SI) was calculated by dividing the percentage of CFSE^lo cells in HBsAg-stimulated wells by the percentage in non-stimulated wells. Values below 1% CFSE^lo cells in non-stimulated wells were set to 1, to exclude unrealistic high values in SI due to division by values <1. The CD4+ T-cell response was considered positive if the SI was >2 and percent CFSE^lo cells in response to HBsAg was at least 5% above the background in non-stimulated wells.

Gordon D., Kelley P., Heinzel S., Cooper P, & Petrovsky N. (2014). Immunogenicity and safety of Advax™, a novel polysaccharide adjuvant based on delta inulin, when formulated with hepatitis B surface antigen; a randomized controlled Phase 1 study. Vaccine, 32(48), 6469-6477.

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Epstein-Barr Virus Immortalization of Lymphocytes

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Blood samples were collected in BD Vacutainer CPT Cell Preparation Tubes containing sodium heparin (Le Pont-de-Claix, France). Peripheral blood mononuclear cells (PBMCs) were collected and processed according to the manufacturer's instructions, and lymphocytes were immortalized by infection with Epstein–Barr virus (EBV) as described [36 (link)]. Briefly, cell lines were established from freshly isolated or cryopreserved lymphocytes using standard EBV transformation protocols which include cell separation by gradient centrifugation and lymphocyte growth enhancement by 1% (v/v) of the mitogenic phytohemagglutinin-M (PHA-M, ThermoFisher Scientific, 10576015). Cell lines were maintained in RPMI 1640 medium supplemented with 20% fetal bovine serum, 2% l-glutamine, 20 units/ml penicillin and 20 µg/ml streptomycin in T75 flasks in 5% CO₂ at 37°C. Cells were maintained at a density of 10⁶ cells/ml, with cell density monitored every other day using trypan blue staining.

Fernández B., Lara Ordóñez A.J., Fdez E., Mutez E., Comptdaer T., Leghay C., Kreisler A., Simonin C., Vandewynckel L., Defebvre L., Destée A., Bleuse S., Taymans J.M., Chartier-Harlin M.C, & Hilfiker S. (2019). Centrosomal cohesion deficits as cellular biomarker in lymphoblastoid cell lines from LRRK2 Parkinson's disease patients. Biochemical Journal, 476(19), 2797-2813.

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Blood Fractionation for PBMC Isolation

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8ml whole blood was collected in BD Vacutainer^® CPT^™ Cell Preparation Tubes and processed within two hours according to manufactures guidelines [19 ]. Briefly, tubes were centrifuged at 1500g for 20 min at 22°C (with no brake) to fractionate the cells using gradient density centrifugation. The cell layer was washed twice with cold phosphate buffered saline before the PBMC pellet was snap frozen on dry ice and transferred to a -80°C freezer until analysis.

Hensman Moss D.J., Robertson N., Farmer R., Scahill R.I., Haider S., Tessari M.A., Flynn G., Fischer D.F., Wild E.J., Macdonald D, & Tabrizi S.J. (2017). Quantification of huntingtin protein species in Huntington’s disease patient leukocytes using optimised electrochemiluminescence immunoassays. PLoS ONE, 12(12), e0189891.

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Isolation and Analysis of Treg and Th17 Cells from Peripheral Blood

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From the peripheral blood of the patients, mononuclear cells (PBMCs) were isolated with BD Vacutainer CPT cell preparation tubes (BD Biosciences, Heidelberg, Germany) according to the manufacturer’s instructions. Immediately after isolation of PBMCs, staining of Treg and Th17 cells was performed with the human Th17/Treg phenotyping Kit (BD Pharmingen, Heidelberg, Germany) according to manufacturer’s staining protocol. Briefly, cells were washed with PBS and stained with markers against CD4, IL-17, and FoxP3 (PerCP-Cy5.5-CD4, PE-IL17 and Alexa Fluor^® 647-FoxP3). Expression of cell surface or intracellular markers was assessed using a flow cytometer (FACS Canto II, Becton Dickinson, Heidelberg, Germany). A typical dot plot and the gating strategy are shown in Figure 4. The frequency of cells related to the total number of CD4⁺ cells was analyzed with FlowJo software: CD4⁺FoxP3⁺ cells were classified as Treg cells and CD4⁺IL17⁺ cells as Th17 cells.

Cucu A., Shreder K., Kraft D., Rühle P.F., Klein G., Thiel G., Frey B., Gaipl U.S, & Fournier C. (2017). Decrease of Markers Related to Bone Erosion in Serum of Patients with Musculoskeletal Disorders after Serial Low-Dose Radon Spa Therapy. Frontiers in Immunology, 8, 882.

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Cryopreservation of Human PBMCs

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Whole-blood samples were collected in BD Vacutainer CPT cell preparation tubes containing sodium heparin as the anti-coagulant (BD, Franklin Lakes, NJ, USA). The CPT tubes were processed within 2 h on at room temperature according to the manufacturer’s instruction. The purified PBMC cells were aliquoted in 90% FBS + 10% dimethylsulphoxide (DMSO) with cell concentration at >2 × 10⁷ cells/ml per cryovial (0.75 ml per cryovial). To ensure stepwise temperature decrease^{37 (link)}, the cryovial were placed in Mr Frosty’ containers (Nalgene/Thermo Fisher, Rochester, NY, USA) with isopropyl alcohol medium in −80 °C freezers for 24–48 h. The cryovials were transferred quickly into liquid nitrogen for long term storage. The crypreserved PBMC samples were shipped with a dry-ice package to the Roswell Park Cancer Center and stored at −8 °C for flow sort sub-cell separation and RNA extraction within 1–2 weeks.

Kuan P.F., Yang X., Clouston S., Ren X., Kotov R., Waszczuk M., Singh P.K., Glenn S.T., Gomez E.C., Wang J., Bromet E, & Luft B.J. (2019). Cell type-specific gene expression patterns associated with posttraumatic stress disorder in World Trade Center responders. Translational Psychiatry, 9, 1.

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Neonatal Endothelial Cell Isolation and Characterization

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Neonatal human dermal blood microvascular ECs (HMVEC), pooled HUVEC, growth medium and supplements were purchased from Lonza (Walkersville, MD). ECs were cultured at 37°C and 5% CO₂. For all experiments, unless otherwise noted, HMVEC were grown on 10 μg/ml collagen until confluent for at least 24 h. For all biochemical experiments at least 10⁴ mAb-coated beads were added per cm². For all magnetic tweezer experiments at least 10³ beads were added per cm². Static force was applied by placing a ceramic magnet above the tissue culture dish for the specified length of time. Primary human neutrophils were isolated from donor blood drawn by BD Vacutainer ® CPT Cell Preparation Tubes (BD Biosciences) following the manufacture’s protocol. Briefly, whole blood was spun for 15 min at 15,000 g. The granulocyte and red blood cell fractions were recovered and the RBCs lysed. The remaining neutrophils were re-suspended in complete EC medium containing 1% Human serum albumin and 1 mM HEPES. Institutional Review Board for University of North Carolina at Chapel Hill has approved all human subject protocols.

Lessey-Morillon E.C., Osborne L.D., Monaghan-Benson E., Guilluy C., O’Brien E.T., Superfine R, & Burridge K. (2014). The RhoA GEF, LARG, mediates ICAM-1-dependent mechanotransduction in endothelial cells to stimulate transendothelial migration. Journal of immunology (Baltimore, Md. : 1950), 192(7), 3390-3398.

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