The anticoagulant activity (anti-Xa activity) of UFH and LMWH was measured in platelet-poor plasma (PPP) at the Comparative Coagulation Laboratory at Cornell University. The PPP was harvested from the supernatant of PRP after high-speed centrifugation (16,000 × g, Accuspin Micro, ThermoScientific, Rockford, IL, USA) for 5 min. The PPP was frozen at −20°C in a dedicated freezer and assays were performed in batch after each treatment for each group of horses with the other investigators remaining blinded as to the results. The PPP was thawed at 37°C in a water bath before analysis. The assay is configured with a bovine activated Factor X reagent added in excess to the test plasma and a chromogenic substrate of Factor Xa (Liquid anti-Xa, Diagnostica Stago, Parsipanny, NJ, USA). The assay is performed using the manufacturer’s automated coagulation analyzer (STA Compact, Diagnostica Stago). In this assay, residual uninhibited Factor Xa cleaves the chromogenic substrate so the inverse of the color change in the reaction mixture is proportional to the drug concentration in the test plasma. Results are expressed as U/mL anti-Xa, based on assay calibration with a standard containing known UFH or LMWH concentrations (STA-calibrators HBPM or LMWH, Diagnostica Stago).
Accuspin micro
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Accuspin Micro is a compact, high-speed microcentrifuge designed for a variety of laboratory applications. It features a brushless DC motor and electronic speed control to provide reliable and consistent performance. The microcentrifuge can achieve a maximum speed of 13,000 rpm and can accommodate rotor capacities up to 24 x 1.5/2.0 mL microtubes.
Automatically generated - may contain errors
Lab products found in correlation
Sta compact, by Diagnostica Stago (2 mentions)
Liquid anti xa, by Diagnostica Stago (1 mentions)
Sta liquid anti xa, by Diagnostica Stago (1 mentions)
Sta apixaban calibrator, by Diagnostica Stago (1 mentions)
Labspec 5, by Horiba (1 mentions)
Branson 2510 ultrasonic cleaner, by Emerson (1 mentions)
4 protocols using accuspin micro
1
Measuring Anticoagulant Activity in Plasma
2
Apixaban Anti-Xa Activity Measurement Protocol
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The anti-Xa activity of apixaban was measured in citrate-anticoagulated PPP by the Comparative Coagulation Laboratory in the Animal Health Diagnostic Center at Cornell University. The PPP was harvested from the supernatant of PRP after high-speed centrifugation (13,000 × g, Accuspin Micro, Thermo Scientific, Rockford, IL, USA) for 5 min. The PPP was frozen at −20°C and assays were performed in batches. The PPP was thawed at 37°C in a water bath before analysis with an automated coagulation analyzer (STA® Compact, Diagnostica Stago, Parsipanny, NJ, USA). The assay is configured with a bovine activated Factor X reagent added in excess to the test plasma and a chromogenic substrate of Factor Xa (STA®-Liquid anti-Xa, Diagnostica Stago). In this assay, residual, uninhibited Factor Xa cleaves the chromogenic substrate such that the inverse of the color change in the reaction mixture is proportional to the drug concentration in the test plasma. Results are expressed as ng/mL anti-Xa activity, based on the calibration standard containing known apixaban concentrations in human plasma (STA®-Apixaban Calibrator, Diagnostica Stago). Assay controls, consisting of apixaban spiked-human plasma (STA®-Apixaban Controls) were measured before each batch of test samples.
3
Raman Spectroscopic Analysis of Copaxone-HA Aggregate
4
Phase Transfer of Oleate-Capped Nanoparticles
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The phase transfer protocol was adapted from
the method developed by Colvin et al.5 (link) Typically,
1 mL of NPs (0.2 mmol) dispersion in chloroform (i.e., stock solution
of the oleate-capped NPs) was added to 500 mg PEG-Monooleate dissolved
in 9 mL chloroform. The solution was stirred overnight at room temperature.
Then, chloroform was removed slowly under vacuum at room temperature,
leaving a waxy layer at the bottom of the flask. About 15 mL of milli-Q
water (18 MΩ) was added to the waxy liquid and dispersed very
well by sonication (2510 Branson Ultrasonic Cleaner) for 30 min. The
murky solution slowly became transparent with sonication. The flask
was put under vacuum and the remaining chloroform was removed slowly
at room temperature. The NPs were collected using a microcentrifuge
(accuSpin Micro, Fisher Scientific) at a speed of 13 000 rpm
for 10 min. The collected pellet was redispersed in milli-Q water
to a concentration of 7.8 mg/mL for cell study. NPs were further diluted
in DPBS to 2000 μg/mL. This stock solution was used for all
subsequent experiments. Nanoparticle stock solutions were ultrasonicated
for 5 min immediately prior to final dilution in culture medium for
cell exposure.
the method developed by Colvin et al.5 (link) Typically,
1 mL of NPs (0.2 mmol) dispersion in chloroform (i.e., stock solution
of the oleate-capped NPs) was added to 500 mg PEG-Monooleate dissolved
in 9 mL chloroform. The solution was stirred overnight at room temperature.
Then, chloroform was removed slowly under vacuum at room temperature,
leaving a waxy layer at the bottom of the flask. About 15 mL of milli-Q
water (18 MΩ) was added to the waxy liquid and dispersed very
well by sonication (2510 Branson Ultrasonic Cleaner) for 30 min. The
murky solution slowly became transparent with sonication. The flask
was put under vacuum and the remaining chloroform was removed slowly
at room temperature. The NPs were collected using a microcentrifuge
(accuSpin Micro, Fisher Scientific) at a speed of 13 000 rpm
for 10 min. The collected pellet was redispersed in milli-Q water
to a concentration of 7.8 mg/mL for cell study. NPs were further diluted
in DPBS to 2000 μg/mL. This stock solution was used for all
subsequent experiments. Nanoparticle stock solutions were ultrasonicated
for 5 min immediately prior to final dilution in culture medium for
cell exposure.
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