Biliverdin IXα was purchased from Frontier Scientific Inc., Logan, UT, spinach ferredoxin-NADP+ reductase, spinach ferredoxin, NADP+, glucose-6-phosphate, and glucose-6-phosphate dehydrogenase were purchased from Sigma-Aldrich. Hemin was obtained from Fluka. Manduca sexta insecticyanin was a gift of Dr. J. Winzerling, University of Arizona. Biliverdin IXγ was obtained by extracting the insecticyanin sample with methanol and removing the precipitated protein by centrifugation. Human factor Xa, prothrombin and thrombin were obtained from Calbiochem. Factor Va was obtained from Haematologic Technologies, and S-2239 was obtained from Diapharma.
Factor va
Manufactured by Prolytix
Factor Va is a laboratory product used in coagulation assays. It is a cofactor required for the activation of prothrombin to thrombin by the prothrombinase complex.
Automatically generated - may contain errors
Lab products found in correlation
Prothrombin, by Prolytix (2 mentions)
Spinach ferredoxin, by Merck Group (1 mentions)
Biliverdin ixα, by Frontier Scientific (1 mentions)
Spinach ferredoxin nadp reductase, by Merck Group (1 mentions)
Glucose 6 phosphate dehydrogenase, by Merck Group (1 mentions)
Argatroban, by Santa Cruz Biotechnology (1 mentions)
Thrombin, by Prolytix (1 mentions)
Factor xa, by Prolytix (1 mentions)
3 protocols using factor va
1
Biliverdin IXα Extraction and Enzymatic Assays
2
Coagulation Factors and Inhibitor Screening
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Factor Va, Xa, prothrombin, and thrombin were obtained from Haematologic Technologies (Essex Junction, VT). The aPTT assay reagent and PT reagents were purchased from Fisher Diagnostics (Middletown, VA). Argatroban was purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX). Inhibitors predicted by high-throughput virtual screening were purchased from ChemBridge (San Diego, CA). All other reagents were of the highest commercially available grade.
3
Microparticle-Stimulated Thrombin Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Microparticle prothrombotic activity was measured in a microparticle capture assay using published methods.30 We treated MΦMP samples with the coagulation factor inhibitors Phe‐Pro‐Arg‐chloromethylketone (50 µmol/L) and Glu‐Gly‐Arg‐chloromethylketone (50 µmol/L), and a 50‐µL microparticle aliquot was added to wells of an annexin V–coated 96‐well plate (StreptaWell plate; Roche, San Francisco, CA; biotinylated annexin V, 0.36 ng/µL coating for 30 minutes; Biovision, Milpitas, CA). After 30‐minute incubation and 3 wash steps, we added prothrombin (1.3 µmol/L), factor Va (2.5 nmol/L) and factor Xa (2.5 nmol/L) (Haematologic Technologies, Inc, Essex Junction, VT) in calcium‐containing Tris buffer (25 mmol/L Tris, 2.5 mmol/L calcium) to the microparticle‐containing wells. Following 30‐minute incubation at 37°C, EDTA addition (0.1M) halted the prothrombinase reaction, and we added Chromozym TH chromogenic thrombin substrate (0.57 mmol/L, Roche) to quantify thrombin activity. The assay measured microparticle‐stimulated thrombin production in reference to a standard curve in a multiplate reader (405 nm optic diameter at 1 minute).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!