Two weeks following the end of stress or an equivalent time frame in non-stressed controls (approximately 29 days from initial handling) rats were injected with 7.5mg/kg MA i.p. and returned to their home cage (CON n=6; CUS n=6). After 30 minutes, the time point with peak efflux in extracellular dorsal striatum dopamine (Figure 2a ), rats were decapitated, whole brains were taken and immediately frozen on dry ice while trunk blood was collected (Vacuette Heparin Tubes, Greiner Bio-One; Radnor, PA). The blood was kept on wet ice until centrifuged at 2000g for 15 min, after which the plasma was pipetted off and stored at −20°C until analysis, while brains were stored at −80°C.
Vacuette heparin tubes
Manufactured by Greiner
Vacuette Heparin Tubes are blood collection tubes designed for the collection and transportation of blood samples. They contain heparin, an anticoagulant, to prevent the blood from clotting during the collection and handling process.
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Lab products found in correlation
2 protocols using vacuette heparin tubes
1
Methamphetamine Effects on Dopamine
2
Isolation of Polymorphonuclear Leukocytes from Whole Blood
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Isolation of cPMNs was performed as previously described [36 (link)]. Venous blood (2 × 10 mL) from controls (n = 13) and periodontitis patients (n = 9) was obtained in lithium heparin tubes (Vacuette® Heparin tubes, Greiner Bio-One, Alphen a/d Rijn, The Netherlands). Blood was diluted 1 : 1 in 1% PBS citrate (pH 7.4). Subsequently, 25 mL of the diluted blood was carefully layered on top of 15 mL Lymphoprep (Axis-shield PoC AS, Oslo, Norway). After centrifugation (800 RCF, 30 min, RT, no brake), the supernatant above the red cell layer was discarded, after which remaining erythrocytes were lysed in cold lysis buffer (NH4Cl (1.5 M), NaHCO3 (100 mM), disodium EDTA (1 mM), all Sigma-Aldrich, Merck, Darmstadt, Germany, 10x diluted in sterile Milli-Q (MQ) water). Immediately after erythrocyte lysis, the cPMN pellet was washed twice in cold PBS (Gibco, Thermo Fisher Scientific, Paisley, Scotland, UK) and recovered in a culture medium (phenol red-free, Roswell Park Memorial Institute (RPMI) 1640, Gibco). All samples were handled on the same day without delay.
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