Platelet-poor plasma (PPP) was obtained from Taipei Blood Center. The plates were incubated with 1 ml PPP at 37 °C for 1 h. Then, 50 μL of the PPP was added into the tube followed by adding 50 μL APTT reagent; the mixture was left to stand for 3 min, and 50 μL of CaCl2 solution was added. The APTT was determined using a coagulation analyzer (CA-50, Sysmex, Kobe, Japan).
Ca 50
Manufactured by Sysmex
Sourced in Japan
The CA-50 is a compact fully automated coagulation analyzer designed for routine testing in clinical laboratories. It is capable of performing a wide range of coagulation assays including prothrombin time (PT), activated partial thromboplastin time (APTT), and fibrinogen. The CA-50 provides efficient workflow and reliable results to support clinical decision-making.
Automatically generated - may contain errors
Lab products found in correlation
Test thrombin reagent, by Siemens (2 mentions)
Dade actin activated cephaloplastin reagent, by Siemens (2 mentions)
Thromborel s, by Siemens (2 mentions)
Actin fsl, by Siemens (1 mentions)
Actin fs activated ptt reagent, by Siemens (1 mentions)
Origin 8, by OriginLab (1 mentions)
Imarktm microplate absorbance reader, by Bio-Rad (1 mentions)
Arvo x5, by PerkinElmer (1 mentions)
Thrombocheck aptt sla, by Sysmex (1 mentions)
18 protocols using ca 50
1
Activated Partial Thromboplastin Time Assay
2
Anticoagulant Properties of Scaffold Materials
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The anticoagulant properties of Gel and SBC/Gel were analyzed using the activated partial thromboplastin time (APTT) by an automated coagulation analyzer CA-50 (Sysmex Co., Kobe, Japan). Healthy fresh human blood was collected intravenously in blood collection tubes and added with acid-citrate dextrose maintenance solution (containing 1.33 g of sodium citrate, 0.47 g of citrate, and 3 g of glucose per 100 ml) at a volume ratio of anticoagulant to blood of 1:4. By centrifuging the anticoagulated whole blood at 3,800 rpm for 10 min, removing the supernatant, and repeating the procedure one more time, platelet-poor plasma (PPP) was prepared. In all blood tests, blood sample from the same donor was used. For each test, 1 mg of scaffold material was incubated with 100 μl of plasma for 5 min at 37 °C before 50 μl of prewarmed APTT reagent (Siemens, Germany) was added. After incubating the mixture at 37 °C for 3 min, 50 μl of prewarmed CaCl2 solution (25 mmol·l–1) was added, mixed, and immediately placed in a coagulometer for testing. The images of the PPP were taken both before and after reaction.
3
Evaluating Antithrombogenicity of Heparinized DLSs
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4
Measuring Coagulation Times with Blood Tester
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The in vitro coagulation times, including the activated partial thrombin time (APTT), were measured using a blood coagulation tester (CA-50; Sysmex Corp., Japan). A sample of 1 × 1 cm2 was placed in 50 μL platelet-poor plasma and incubated at 37°C for 1 min, followed by adding 50 μL APTT reagent. After incubation for 3 min, 50 μL CaCl2 was added. The APTT was then determined.
5
Plasma Coagulation Time Measurement
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Each cell line was incubated with Vor, L-asp, or Dox for 8 or 24 h. EAhy926 cells and HepG2 cells were confluent and the other tumor cell lines proliferated in suspension. EAhy926 cells and HepG2 cells were taken off from plates by 0.25% trypsin and washed with phosphate-buffered saline (PBS) twice. A portion of cells (2 × 106) was suspended in 50 µL of PBS, and added to 50 µL of pooled normal human plasma. After incubation at 37°C for 3 min, 50 µL of 25 mM calcium chloride was added and the plasma recalcification time measured using a semi-automatic coagulator (CA-50; Sysmex, Kobe, Japan).9 (link),12 (link)
6
Coagulation Assay: APTT and TT Measurement
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Activated partial thromboplastin time (APTT) and thrombin time (TT) were measured by an automated blood coagulation analyzer CA-50 (Sysmex Corporation, Kobe, Japan), and the test method was described as follows: fresh blood was collected using vacuum tubes, containing sodium citrate as an anticoagulant (anticoagulant to blood ratio, 1:9, v/v). The platelet-poor plasma (PPP) was obtained after centrifuging at 4000 rpm for 15 min. Synchronously, the samples (0.4 cm×0.5 cm) were immersed in PBS (0.2 mL, pH = 7.4) for 1 h. Then the PBS was removed and 0.1 mL of fresh PPP was introduced. After incubating at 37°C for 30 min, 50μL of the incubated PPP was added into the test cup, followed by the addition of 50μL of APTT agent (incubated 10 min before use) and incubation at 37°C for 3 min. Thereafter, 50μL of 0.025 M CaCl 2 solution was added, and then the APTT was measured. For the TT test, 50μL of TT agent was added into the test cup (containing 50μL of the incubated PPP) after 10 min incubating, and then the TT was measured. At least three measurements were averaged to get a reliable value, and the results were analyzed by statistical method.
7
Coagulation Profile of Stents in Platelet-Poor Plasma
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Anticoagulated blood sample was centrifuged at 3000 rpm for 5 min to prepare platelet-poor plasma (PPP). Five different stent groups were incubated in 0.5 mL PPP at 37 °C for 30 min. 50 µl PPP was transferred to a special test tube, then 100 µl APTT reagent (Sichuan Maker Science Technology Co, China), 100 µl PT reagent, 100 µl TT reagent and 100 µl fibrinogen (Fbg) reagent were added into the tube respectively and incubated at 37 °C for 3 min. Finally, the APTT, PT, TT and Fbg content were measured by Automatic Coagulation Analyzer (CA-50, Sysmex, Japan) (n = 3).
8
Coagulation Assays for Anticoagulant Screening
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The APTT and thrombin time (TT) were measured using an automated blood coagulation analyzer (CA-50; Sysmex co, Kobe, Japan). Platelet-poor plasma obtained from sodium citrate-anticoagulated human blood was incubated with vehicle or drugs for 1 minute at 37°C. The APTT reagent (Dade Actin FSL) was added into plasma and incubated for an additional 3 minutes. Finally, CaCl 2 (8.3 mmol/L) was added to initiate the coagulation cascade, and the APTT was determined as the time required to form a fibrin clot. For TT assay, Dade thrombin reagent was used to initiate the coagulation process. The upper limit of detection of APTT or TT was 190 seconds.
9
Plasma Coagulation Assay for Cell-Derived Procoagulant Activity
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Each cell line was treated with IDR, AraC, or Am80 for 8 or 24 h. EAhy926 cells were confluent and the other tumor cell lines proliferated in suspension. EAhy926 cells were collected by treatment with 0.05% trypsin and washed with phosphate-buffered saline (PBS) twice. A portion of the cells (2 × 106) was suspended in 50 μL PBS and combined with 50 μL pooled normal human plasma. Normal blood was collected from healthy individuals using a syringe with 109 mM sodium citrate, a tourniquet, and a 22-G needle. Blood cells were removed from plasma by centrifugation for 15 min at 2,000 g using a refrigerated centrifuge. The resulting supernatant was immediately transferred into a clean polypropylene tube using a Pasteur pipette. The plasma was apportioned into 2 ml aliquots, stored at –80°C. For the PCA assay plasma from five healthy individuals was pooled. We avoided freeze-thaw cycles. After incubation at 37°C for 3 min, 50 μL of 25 mM calcium chloride was added, and the plasma recalcification time was measured using a semi-automatic coagulator (CA-50; Sysmex, Kobe, Japan) [12 (link), 13 (link)]. CA-50 detects a change in plasma optical density during blood coagulation, that is, fibrin formation. Shortening of coagulation time indicates increased PCA.
10
Evaluating Antithrombogenicity of Stimuli-Responsive Hemicellulose Microgels
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To evaluate the antithrombogenicity of
the stimuli-responsive hemicellulose microgels, activated partial
thromboplastin time (APTT) and thrombin time (TT) were measured by
an automated blood coagulation analyzer CA-50 (Sysmex Corporation,
Kobe, Japan) according to the method described in a previous report.41 Healthy human fresh blood was collected in vacuum
tubes containing sodium citrate as the anticoagulant (anticoagulant
to blood ratio, 1:9 v/v), and the platelet-poor plasma (PPP) was obtained
after centrifuging at 4000 rpm for 15 min. Stimuli-responsive hemicellulose
microgels (10 mg/mL) were prepared in buffer solution (pH = 7.4),
5 μL suspensions and 100 μL of PPP were incubated at 37
°C for 0.5 h, and 50 μL of incubated solution was then
added to the test cup, followed by the addition of 50 μL of
APTT agent (Dade Actin Activated Cephaloplastin Reagent, Siemens;
incubated 10 min before use). The solution was incubated at 37 °C
for another 3 min, and 50 μL of 0.025 M CaCl2 was
subsequently added. The APPT was recorded by an automated blood coagulation
analyzer CA-50. Two independent measurements were averaged to reach
a reliable value. The TT test was performed in a process similar to
that of the APTT test. The only difference is the APTT agent replaced
by Test Thrombin Reagent (Siemens; incubated 10 min before use).
the stimuli-responsive hemicellulose microgels, activated partial
thromboplastin time (APTT) and thrombin time (TT) were measured by
an automated blood coagulation analyzer CA-50 (Sysmex Corporation,
Kobe, Japan) according to the method described in a previous report.41 Healthy human fresh blood was collected in vacuum
tubes containing sodium citrate as the anticoagulant (anticoagulant
to blood ratio, 1:9 v/v), and the platelet-poor plasma (PPP) was obtained
after centrifuging at 4000 rpm for 15 min. Stimuli-responsive hemicellulose
microgels (10 mg/mL) were prepared in buffer solution (pH = 7.4),
5 μL suspensions and 100 μL of PPP were incubated at 37
°C for 0.5 h, and 50 μL of incubated solution was then
added to the test cup, followed by the addition of 50 μL of
APTT agent (Dade Actin Activated Cephaloplastin Reagent, Siemens;
incubated 10 min before use). The solution was incubated at 37 °C
for another 3 min, and 50 μL of 0.025 M CaCl2 was
subsequently added. The APPT was recorded by an automated blood coagulation
analyzer CA-50. Two independent measurements were averaged to reach
a reliable value. The TT test was performed in a process similar to
that of the APTT test. The only difference is the APTT agent replaced
by Test Thrombin Reagent (Siemens; incubated 10 min before use).
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