Acid citrate dextrose

Manufactured by Merck Group

Acid citrate dextrose is a solution used in various laboratory procedures. It is composed of citric acid, sodium citrate, and dextrose. The primary function of acid citrate dextrose is to serve as an anticoagulant, preventing the clotting of blood samples during collection and storage.

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Lab products found in correlation

Edta tube, by BD (2 mentions) Hemavet 950fs, by Drew Scientific (2 mentions) 40 μm pore size cell strainer, by BD (1 mentions) Red blood cell lysis buffer, by BioLegend (1 mentions) Balb c mice, by Jackson ImmunoResearch (1 mentions) Addavax adjuvant, by InvivoGen (1 mentions) Facscanto 2, by BD (1 mentions) Rbc lysis buffer, by BD (1 mentions) Lsrii flow cytometer, by BD (1 mentions) Anti ly6c pe, by BD (1 mentions) Fcs express software, by De Novo Software (1 mentions) Anti cd41 allophycocyanin, by Thermo Fisher Scientific (1 mentions) Anti cd11b efluor450, by Thermo Fisher Scientific (1 mentions) Fitc dextran, by Merck Group (1 mentions) Ficoll histopaque 1077, by Merck Group (1 mentions) Centrifuge 5804 r, by Eppendorf (1 mentions) Dulbecco phosphate buffered saline (dpbs), by Thermo Fisher Scientific (1 mentions) Trypan blue staining, by Merck Group (1 mentions) Ammonium chloride, by Merck Group (1 mentions) Potassium bicarbonate, by Merck Group (1 mentions)

Close spelling variants of this product

Dextrose (252 citations) Acid-citrate-dextrose solution (12 citations) Acid-citrate-dextrose buffer (6 citations) Citrate acid (5 citations)

9 protocols using acid citrate dextrose

Antigen-Specific Immune Response Study

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Six-week-old female BALB/c mice were purchased from The Jackson Laboratory. The mice were housed in ventilated cages in environmentally controlled rooms at the Scripps Research Institute (TSRI), in compliance with an approved IACUC protocol and AAALAC guidelines. At week 0, each mouse was immunized with 100 μl (50 μg) of antigen formulated in 50 μl AddaVax adjuvant (InvivoGen) per the instructions of the manufacturer via the subcutaneous route. At week 3 and week 6, the animals were boosted with 10 μg of antigen formulated in AddaVax adjuvant. At week 8, the animals were terminally bled by cardiac puncture and the anticoagulant acid citrate dextrose (Sigma-Aldrich) was added to the samples at a 1:10 ratio. Samples were spun at 1,000 rpm for 10 min at 4°C to separate plasma and cells. Red blood cell lysis buffer (BioLegend) was added to the cell fraction. After 2 rounds of washing with PBS, peripheral blood mononuclear cells (PBMCs) were resuspended in Bambanker freezing media (Lymphotec Inc.). Spleens were also harvested and ground against a 40-μm-pore-size cell strainer (BD Falcon) to release the splenocytes into a cell suspension. The cells were centrifuged, treated with 10 ml of red blood cell (RBC) lysis buffer per manufacturer specifications, and resuspended in Bambanker freezing media for cell freezing.

Morris C.D., Azadnia P., de Val N., Vora N., Honda A., Giang E., Saye-Francisco K., Cheng Y., Lin X., Mann C.J., Tang J., Sok D., Burton D.R., Law M., Ward A.B., He L, & Zhu J. (2017). Differential Antibody Responses to Conserved HIV-1 Neutralizing Epitopes in the Context of Multivalent Scaffolds and Native-Like gp140 Trimers. mBio, 8(1), e00036-17.

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Resolvin D3 Modulates PMN-Platelet Interactions

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Fresh human blood was collected in acid citrate–dextrose (Sigma-Aldrich) from healthy volunteers giving informed consent under protocol # 1999-P-001297 approved by the Partners Human Research Committee. Immediately after collection, 500 µL of whole blood were incubated with RvD3 (1–100 nM) or vehicle (0.05% EtOH in PBS) for 15 min at 37°C, followed by addition of PAF (500 nM) or vehicle (0.05% EtOH in PBS) and incubations continued for 20 min. To assess PMN-platelet interactions samples were stained with mouse anti-human CD16-PE and mouse anti-human CD41-FITC (20 min, RT), red blood cells lysed (RBC lysis buffer (BD); 10 min) and cells washed. Staining was assessed immediately using FACS Canto II (BD Biosciences) and analyzed using FlowJo (TreeStar).

Norris P.C., Arnardottir H., Sanger J.M., Fichtner D., Keyes G.S, & Serhan C.N. (2016). Resolvin D3 multi-level proresolving actions are host protective during infection. Prostaglandins, leukotrienes, and essential fatty acids, 138, 81-89.

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Murine Leukocyte and Platelet Profiling

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Whole blood samples from mice were collected and maintained in acid citrate dextrose (Sigma-Aldrich; Merck KGaA). Anti-CD11b eFluor450 (1:100), anti-CD41-allophycocyanin (1:100) (both from eBioscience; Thermo Fisher Scientific, Inc.), anti-Ly6C-PE (1:100; BD Biosciences, Franklin Lakes, NJ, USA) and anti-Ly6G-phycoerythrin (1:100; BD Biosciences) were used to detect leukocyte and platelets antigens. Samples were examined with an LSRII flow cytometer (BD Biosciences) and analyzed using FCS express software (version 3.0, De Novo Software, Glendale, CA, USA). Neutrophils and monocytes were gated through their forward- and side-scatter characteristics and through their Ly-6G⁺/CD11b⁺ (neutrophil) and Ly-6G⁻/CD11b⁺/Ly-6C⁺ (monocyte) expression pattern. Platelet-neutrophil and platelet-monocyte aggregates were calculated using CD41 antibody staining.

Han X., Wu Y.C., Meng M., Sun Q.S., Gao S.M, & Sun H. (2018). Linarin prevents LPS-induced acute lung injury by suppressing oxidative stress and inflammation via inhibition of TXNIP/NLRP3 and NF-κB pathways. International Journal of Molecular Medicine, 42(3), 1460-1472.

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Quantifying Intestinal Permeability Using FITC-Dextran

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FITC-dextran (70 kilodaltons) and acid-citrate-dextrose solutions were purchased from Sigma-Aldrich. Mice were fasted for 6 hours before the test. Then, mice were administered a dose (150 uL/mouse) of FITC-dextran in sterile 1× PBS (80 mg/mL) through gavage technique. After 4 hours, blood was collected in a capillary tube by nicking the tail of the mouse in its proximal region. acid-citrate-dextrose solution was added 15% vol/vol as an anticoagulant. Then, all blood samples were centrifuged at 5000 rpm for 10 minutes. Plasma was collected in a new 1.5-mL tube and kept in the dark at 4°C. Plasma then was diluted 1:10 in PBS and transferred to a black, opaque-bottom, 96-well plate. A volume of 100 uL was transferred into each well. PBS was used as blank. Readings of RFUs were measured by a SpectraMax i3x (San Jose, CA) spectrophotometer in triplicate and averaged. Fluorescence was determined at 530 nm with excitation at 485 nm.

Di Martino L., De Salvo C., Buela K.A., Hager C., Ghannoum M., Osme A., Buttò L., Bamias G., Pizarro T.T, & Cominelli F. (2021). Candida tropicalis Infection Modulates the Gut Microbiome and Confers Enhanced Susceptibility to Colitis in Mice. Cellular and Molecular Gastroenterology and Hepatology, 13(3), 901-923.

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Isolation of Human Neutrophils

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Whole blood samples were drawn from cubital veins from healthy donors and anticoagulated with 1 ml acid–citrate-dextrose (Sigma-Aldrich) per 5 ml whole blood according to approved Institutional Review Board (University of Illinois at Chicago, Chicago, IL) protocol. Samples were suspended in a three-fold volume of ice-cold lysis medium containing 155 mM of ammonium chloride and 10 mM of potassium bicarbonate (both from Sigma-Aldrich) and kept on ice for 10 min. After brief centrifugation at 1,500 rpm and 4°C (Eppendorf Centrifuge 5804 R; Eppendorf Hamburg, Germany), the supernatant was discarded and the cell pellet was resuspended in 2 ml of ice-cold lysis medium supplemented with 0.5% bovine serum albumin. The suspension was then kept on ice 15 min followed by another centrifugation at 1,500 rpm and 4°C. Again, the supernatant was discarded. The pellet was resuspended in 2 ml of ice-cold Dulbecco phosphate-buffered saline (Gibco Invitrogen) and layered on 2 ml of Ficoll Histopaque 1077 (Sigma-Aldrich). After 30 min of centrifugation at 1,500 rpm and 4°C with low brake, the supernatant was discarded and the pellet consisting of neutrophils was resuspended in Hank balanced salt solution (Gibco Invitrogen). Viability was 95% or more as assessed with Trypan blue staining (Sigma-Aldrich).

Piegeler T., Votta-Velis E.G., Bakhshi F.R., Mao M., Carnegie G., Bonini M.G., Schwartz D.E., Borgeat A., Beck-Schimmer B, & Minshall R.D. (2014). Endothelial Barrier Protection by Local Anesthetics: Ropivacaine and Lidocaine Block Tumor Necrosis Factor-α–induced Endothelial Cell Src Activation. Anesthesiology, 120(6), 1414-1428.

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Whole Blood Analysis Protocol

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For whole blood analysis, blood was withdrawn from the submandibular vein for survival bleeding or by cardiac puncture in terminal experiments. Blood was collected into either acid citrate dextrose (Sigma-Aldrich) for flow cytometry analysis or EDTA tubes (BD microtainer) for platelet counting. Platelet counts in the peripheral blood were measured with a Hemavet 950 FS complete blood counting instrument (Drew Scientific) and confirmed with manual platelet counts.

Lefrançais E., Ortiz-Muñoz G., Caudrillier A., Mallavia B., Liu F., Sayah D.M., Thornton E.E., Headley M.B., David T., Coughlin S.R., Krummel M.F., Leavitt A.D., Passegué E, & Looney M.R. (2017). The lung is a site of platelet biogenesis and a reservoir for hematopoietic progenitors. Nature, 544(7648), 105-109.

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Whole Blood Analysis Protocol

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Platelet Enumeration from Blood Samples

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To determine platelet number, 18 µl of blood extracted from the submandibular vein was mixed with 2 µl acid citrate dextrose (Sigma-Aldrich). Ten microliters of the mixture was then blended with 190 µl NaNH₃ (Sigma-Aldrich) for 5 min at RT to induce the lysis of red blood cells. Then, 10 µl of this solution was diluted in 90 µl of phosphate-buffered saline (PBS). Counts were performed in a Burker chamber on a microscope with a 40× objective (Dhanjal et al., 2007 (link)).

Cordero-Sanchez C., Riva B., Reano S., Clemente N., Zaggia I., Ruffinatti F.A., Potenzieri A., Pirali T., Raffa S., Sangaletti S., Colombo M.P., Bertoni A., Garibaldi M., Filigheddu N, & Genazzani A.A. (2019). A luminal EF-hand mutation in STIM1 in mice causes the clinical hallmarks of tubular aggregate myopathy. Disease Models & Mechanisms, 13(2), dmm041111.

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Isolation and Purification of CD34+ Hematopoietic Progenitor Cells

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Adult peripheral blood was extracted by injecting approximately 50 ml of Dulbecco’s PBS (DPBS) with 10% acid citrate dextrose (Sigma Aldrich) into leukocyte reduction system (LRS) chambers discarded following plateletpheresis from two male subjects and allowing the chamber to drain under gravity. Umbilical cord blood units obtained from one male and one female donor, were diluted in a 1:1 ratio with DPBS. Mononuclear cells from both adult and paediatric samples were then obtained by density gradient centrifugation (Ficoll-Paque PLUS, GE Healthcare). Positive selection of CD34+ haematopoietic progenitor cells was then performed using CD34 MicroBead Kit Ultrapure Microbeads (Miltenyi Biotec) at a volume of 50 µl per 10⁸ cells, with an autoMACS Pro Separator (Miltenyi Biotec) as per manufacturer’s instructions. Purified cells were plated at a density of 5 × 10⁴ cells/ml of media.

Ong L.T., Schibeci S.D., Fewings N.L., Booth D.R, & Parnell G.P. (2021). Age-dependent VDR peak DNA methylation as a mechanism for latitude-dependent multiple sclerosis risk. Epigenetics & Chromatin, 14, 9.

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