Platelet aggregation was performed on a Chronolog aggregometer (Chrono-Log Corp., PA, USA) as previously described [31 (link),32 (link)]. Briefly, fresh PRP prepared from subjects was stimulated by ADP or collagen at baseline, at 6 weeks, and at 12 weeks. Platelet aggregation was evaluated on a Chronolog aggregometer (Chrono-Log Corp., PA, USA) in PRP (3.0 × 108 platelets/mL) at 37 °C with a sample stir speed of 1000 rpm, and the change in light transmission was monitored and recorded for at least 6 minutes.
Aggregometer
Manufactured by Chrono-Log
Sourced in United States
The Aggregometer is a laboratory instrument used to measure the aggregation of platelets in blood samples. It detects and quantifies the clumping of platelets in response to various agonists. The Aggregometer provides real-time monitoring of platelet function, enabling researchers to assess the effectiveness of antiplatelet drugs or investigate platelet disorders.
Automatically generated - may contain errors
Lab products found in correlation
Thrombin, by Merck Group (6 mentions)
Adenosine diphosphate (adp), by Chrono-Log (2 mentions)
Luciferin luciferase kit, by Chrono-Log (1 mentions)
Axiovert 200m, by Zeiss (1 mentions)
Multiplate platelet function analyzer, by Roche (1 mentions)
Su8220, by Hitachi (1 mentions)
U46619, by Chrono-Log (1 mentions)
Synergy ht multi model microplate reader, by Agilent Technologies (1 mentions)
Bovine thrombin, by Merck Group (1 mentions)
Optical aggregometer, by Chrono-Log (1 mentions)
Lsrfortessa flow cytometer, by BD (1 mentions)
Buffered sodium citrate, by BD (1 mentions)
Animal blood counter, by Horiba (1 mentions)
Fibrillar type 1 collagen, by Takeda Pharmaceuticals (1 mentions)
Aggrolink 8, by Chrono-Log (1 mentions)
U46619, by Cayman Chemical (1 mentions)
Ht1002a, by Stago (1 mentions)
Scil vet abc plus, by Horiba (1 mentions)
Cytexpert 2, by Beckman Coulter (1 mentions)
Cytoflex, by Beckman Coulter (1 mentions)
102 protocols using aggregometer
1
Platelet Aggregation Analysis Protocol
2
Evaluating Anti-Platelet Aggregation
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Example 12
One hour following p.o. dosing with a test compound whole blood is collected from male beagle dogs in a 5 mL Vacutainer with exogenous heparin (5 U/mL) added to the Vacutainer. Aggregation studies are evaluated by using a whole blood Aggregometer (Chronolog Corp.). Briefly, whole blood (400 μL) is added to saline (600 μL) with constant stirring and activated with 5 μg of Collagen (Chronolog Corp.). The serotonin response is obtained by adding 5-HT (Sigma) to a final concentration of 2.5 μM.
Results:
Selected compounds are tested for anti-platelet aggregation activity after single bolus oral dosing. The dose that affords maximal inhibition of 5-HT amplified platelet aggregation is identified and used for comparison.
3
Ex-Vivo Canine Platelet Aggregation
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Example 12
Ex-Vivo Dog Whole Blood Aggregation
One hour following PO dosing with a test compound whole blood was collected from male beagle dogs in a 5 mL vacutainer with exogenous heparin (5 U/mL) added to vacutainer. Aggregation studies were evaluated by using whole blood Aggregometer (Chronolog Corp.). Briefly, whole blood (400 uL) was added to saline (600 uL) with constant stirring and activated with 5 ug of Collagen (Chronolog Corp.). The serotonin response was obtained by adding 5-HT (Sigma) to final concentration of 2.5 μM. Results: Selected compounds were tested for antiplatelet aggregation activity after single bolus oral dosing. The dose that afforded maximal inhibition of 5-HT amplified platelet aggregation was identified and used for comparison.
4
Quantification of Platelet Secretion
5
Platelets Aggregation Assay with G-Ro
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Washed human platelets (108/mL) were preincubated with or without G-Ro in the reaction system containing 2mM of CaCl2 for 3 min at 37°C, then stimulated with thrombin (0.05 U/mL). The aggregation was performed for 5 min using an aggregometer (Chrono-Log Corporation, Havertown, PA, USA) at a constant stirring speed of 1,000 rpm. Each aggregation rate was determined as an increase in light transmission. G-Ro was dissolved in platelet suspension buffer (pH 6.9), and suspension buffer was used as the reference (transmission 0)
6
Platelet Aggregation Monitoring Protocol
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Platelet aggregation was monitored by measuring light transmission through the stirred suspension of washed platelets (3 × 108/ml) at 37°C using a Chronolog aggregometer. Platelet aggregation was triggered by adding fibrillar collagen, convulxin, ADP, or thrombin. Representative traces for aggregation were obtained from at least three independent experiments.
7
Platelet Aggregation Assay Protocol
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The in vitro platelet aggregation study was performed according to a previously reported method 17 , 18 . Platelet‐rich plasma (PRP) was incubated with the indicated concentration of each compound in DMSO for 1, 3, 5 or 10 min. They were subsequently stimulated by U46619 (2 μM) in 0.9% saline solution or collagen (1 μg/ml) at 37°C for 5 min. Platelet aggregation was recorded using an aggregometer (Chronolog, Havertown, PA, USA). For the ex vivo aggregation assay, male mice were fasted overnight and the indicated concentration of each compound in DMSO was administered by intravenous (i.v.) injection. After 24 hrs, PRP (109 platelets/ml) in a volume of 240 μl was incubated at 37°C for 1.5 min. in the aggregometer under continuous stirring at 1000 r.p.m. and subsequently stimulated with U46619 (2 μM). Platelet aggregation was recorded as described above.
8
Platelet Aggregation and Spike Protein
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Platelets (2.5 × 108 platelets per mL) were incubated with 2 μg/mL spike protein for 10 min, then stimulated with different agonists. To explore the effect of CoQ10 on spike protein-potentiated platelet aggregation, gel-filtered platelets were pre-incubated with 100 μM CoQ10 or solvent (0.5% DMSO) in the dark, then incubated with 2 μg/mL spike protein for 10 min and platelet aggregation was stimulated with 0.1 U/mL thrombin. Platelet aggregation was evaluated by an aggregometer (Chrono-Log Corp.). The concentration of CoQ10 is referred to our previous study [11 (link)]. For platelet activation assessment, platelets were incubated with FITC-conjugated anti-human CD62P or PAC-1 antibodies, or Alexa FluorTM 488-conjugated fibrinogen for 20 min at room temperature after pre-treatment. Platelets were then stimulated with 0.1 U/mL thrombin for 10 min or not, and were fixed with 1% paraformaldehyde and then analyzed using flow cytometry (Backman Coulter Inc., Pasadena, CA, USA).
9
Platelet Aggregation Assay with Compounds
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The platelet aggregation assay was measured following to the previously reported method [44 (link)]. Washed plasma was pre-incubated with the indicated concentration of compounds 5 , 9 , and 21 –23 for 3 min, and then stimulated with thrombin (0.1 U/mL, Sigma) and U46619 (2 µM) in 0.9% saline for 5 min. Platelet aggregations were performed in an aggregometer (Chronolog, Havertown, PA, USA).
10
Kindlin-3 Phosphorylation and Platelet Aggregation
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Effect of thrombin or PMA on kindlin-3 phosphorylation was examined using a kindlin-3 phosphospecific mAb while monitoring aggregation in an aggregometer (Chrono-log Corporation). Platelets (3 × 108/ml) were equilibrated at 37°C for 5 min. Then, an agonist was added and aggregation was monitored for times indicated at 37°C with stirring. The final volume in each cuvette was 500 μl. Platelets were solubilized in a Laemmli buffer containing 62.5 mM Tris–HCl, pH 7.4, 2% SDS, 5% 2-mercaptoethanol, and 10% glycerol and subjected to SDS–PAGE. Proteins were transferred to PVDF, and blots probed with antibodies of interest using 5% BSA as a blocking agent.
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