The following coagulation and coagulation-related proteases were purchased from Enzyme Research Labs (USA): kallikrein and thrombin [also called factor (F) IIa, FXa, FXIa, and FXIIa]. The following chromogenic substrates were purchased from Diapharma (USA): for thrombin, S-2238; for FXa, S-2765; for FXIa and APC, S-2366; and for kallikrein and FXIIa, S-2302. Custom synthetic double-stranded DNA fragments (gBlocks) were bought from Integrated DNA Technologies (Canada). Restriction endonucleases and glutathione agarose were purchased from Thermo Fisher Scientific (Canada). Nickel chelate affinity resin Ni-NTA agarose was bought from Qiagen (Canada). PreScission Protease [a glutathione sulfotransferase (GST)–human rhinovirus (HRV) 3C protease fusion protein] was purchased from GE Healthcare (Canada). Normal human pooled plasma (NHPP) was produced in-house. FXI-deficient plasma was purchased from Affinity Biologicals (Canada). STA PTTA reagent, STA Neoplastine CI Plus reagent, and Owren-Koller buffer were bought from Diagnostica Stago (Canada).
S 2366
Manufactured by Diapharma
Sourced in United States
The S-2366 is a laboratory instrument designed for analytical and research applications. It is capable of measuring and analyzing various parameters of samples. The core function of the S-2366 is to provide accurate and reliable data to support scientific investigations and experiments. Details about the intended use or specific applications of the S-2366 are not available.
Automatically generated - may contain errors
Lab products found in correlation
S 2302, by Diapharma (5 mentions)
Kallikrein, by Diapharma (2 mentions)
Kallikrein, by Enzyme Research (2 mentions)
S 2238, by Diapharma (2 mentions)
S 2765, by Diapharma (2 mentions)
Protac, by Sekisui (2 mentions)
Spectrozyme, by Sekisui (2 mentions)
Glutathione agarose, by Thermo Fisher Scientific (1 mentions)
Restriction endonuclease, by Thermo Fisher Scientific (1 mentions)
Sta ptta reagent, by Diagnostica Stago (1 mentions)
Owren koller buffer, by Diagnostica Stago (1 mentions)
Prescission protease, by GE Healthcare (1 mentions)
Heparin, by Pfizer (1 mentions)
Protamine, by Fresenius (1 mentions)
Chondroitinase abc, by Merck Group (1 mentions)
Heparinase 1 3, by Merck Group (1 mentions)
Hyaluronidase, by Merck Group (1 mentions)
Calf thymus histone, by Merck Group (1 mentions)
Thrombin, by Diapharma (1 mentions)
Thrombin, by Enzyme Research (1 mentions)
12 protocols using s 2366
1
Purification of Coagulation Proteases
2
Assay for Intrinsic Coagulation Factor
3
Activated Protein C Biochemical Assay
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Recombinant human APC (hAPC, drotrecogin alfa [activated]) was purchased from Eli Lilly (Indianapolis, IN, USA); plasma-derived hAPC and hPC from Enzyme Research Laboratories (South Bend, IN, USA); calf thymus histones from Sigma-Aldrich (St. Louis, MO, USA); TM and human coagulation FVa from Hematologic Technologies (Essex Junction, VT, USA); Protac, a rapid activator of PC from Sekisui Diagnostics (Lexington, MA, USA); hAPC substrate Spectrozyme PCa from American Diagnostica (Pfungstadt, Germany) and Sekisui Diagnostics (Lexington, MA, USA), and S-2366 from Diapharma (West Chester, OH, USA).
4
Purified Coagulation Protease Assays
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Purified human coagulation proteases FXIa, thrombin, FXa, FXIIa, kallikrein, and APC were from Enzyme Research Labs (South Bend, IN, USA). Chromogenic substrates were purchased from Diapharma (West Chester, OH, USA): for FXIa and APC, S-2366; for kallikrein and FXIIa, S-2302; for thrombin, S-2238; and for FXa, S-2765.
5
Kinetic Assay and ELISA for C1-INH Activity
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PdC1-INH (Cetor®) was obtained from the Sanquin Blood Supply Foundation, The Netherlands. Monoclonal antibody (mAb) RII and polyclonal antibodies against C1-INH were produced previously [36 (link),37 (link)]. C1s for kinetic assays and ELISA was purchased from Calbiochem (La Jolla, CA, USA). C1s and pAb against C1-INH were biotinylated using EZ-Link® Sulfo-NHS-Biotin (Thermo Fisher Scientific, Waltham, MA, USA) or Pierce (Rockford, ILL) following the manufacturer protocol. Human Factor XIIa and kallikrein were purchased from Kordia (Leiden, The Netherlands). Agarose-bound RCA120 and biotinylated-RCA120 were from Vector Labs, USA. Streptavidin-peroxidase was obtained from Amersham Pharmacia Biotech (Uppsala, Sweden). Streptavidin-polymerized peroxidase (poly-HRP) was obtained from Sanquin (Business Unit Reagents, Amsterdam, The Netherlands). Chromogenic substrates S2251, S-2302 and S2366 were from DiaPharma (Beckett Ridge, Ohio, USA); C1s substrate H-D-Val-Ser-Arg.p-NA.2HCl was custom synthesized by Biomatik (Wilmington, DE, USA).
6
Characterization of Thrombin Binding Interactions
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Enzymes were purchased commercially from Haemtech Biopharma Services, except for those used in the experiments shown in section “Binding of dansyl Compound 2 to thrombin” below, where wild-type thrombin and mutant S195A were expressed as prethrombin-2 in E. coli, refolded, and purified to homogeneity as previously described [17 ]. Chromogenic substrates S-2238, S-2302, S-2366, and S-2765 were purchased from Diapharma, Spectrozyme was purchased from Sekisui Diagnostics, and fibrinogen was purchased from Sigma.
7
Plasma Proteins for Coagulation Assays
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Human plasma proteins
including thrombin, factors Xa, XIa, FXIa-DEGR, and XI were obtained
from Haematologic Technologies (Essex Junction, VT). Stock solutions
of factors XIa, XI, and thrombin were prepared in 50 mM Tris-HCl buffer,
pH 7.4, containing 150 mM NaCl, 0.1% PEG8000, and 0.02% Tween80. Stock
solution of factor Xa was prepared in 20 mM Tris-HCl buffer, pH 7.4,
containing 100 mM NaCl, 2.5 mM CaCl2, 0.1% PEG8000, and
0.02% Tween80. Chromogenic substrates including Spectrozyme TH (H-d -cyclohexylalanyl-Ala-Arg-p-nitroanilide) and Spectrozyme factor Xa (methoxycarbonyl-d -cyclohexylglycyl-Gly-Arg-p-nitroanilide) were obtained
from American Diagnostica (Greenwich, CT). S-2366 (l -PyroGlu-Pro-Arg-p-nitroaniline HCl) was obtained from Diapharma (West Chester,
OH). FXIa-CD was a gift from Dr. Alireza Rezaie of Saint Louis University.
including thrombin, factors Xa, XIa, FXIa-DEGR, and XI were obtained
from Haematologic Technologies (Essex Junction, VT). Stock solutions
of factors XIa, XI, and thrombin were prepared in 50 mM Tris-HCl buffer,
pH 7.4, containing 150 mM NaCl, 0.1% PEG8000, and 0.02% Tween80. Stock
solution of factor Xa was prepared in 20 mM Tris-HCl buffer, pH 7.4,
containing 100 mM NaCl, 2.5 mM CaCl2, 0.1% PEG8000, and
0.02% Tween80. Chromogenic substrates including Spectrozyme TH (H-
from American Diagnostica (Greenwich, CT). S-2366 (
OH). FXIa-CD was a gift from Dr. Alireza Rezaie of Saint Louis University.
8
Coagulation Factors and Inhibitors Protocol
9
Fluorescein-labeled fXIa Enzyme Characterization
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Human factor XIa (fXIa and active-site labeled
fluorescein-EGR-fXIa (fEGR-fXIa)) was purchased from
Haematologic Technologies (Essex Junction, VT). Recombinant fXIa containing
only the catalytic domain (fXIa-CD) was a gift from Dr. Alireza Rezaie
(St. Louis University, MO). Chromogenic substrate S-2366 (l -pyroglutamyl-l -prolyl-l -arginine-p-nitroaniline) was purchased from Diapharma (West Chester, OH). Stock
solutions of fXIa were prepared in 0.05 M Tris-HCl buffer, pH 7.4,
containing 0.15 M NaCl and 0.1% PEG8000. The buffer used in inhibition
studies was 0.05 M Tris-HCl buffer, pH 7.4, containing 0.15 M NaCl,
0.1% PEG8000, and 0.02% Tween80, while that used for all other studies
was devoid of 0.02% Tween80. All of the other chemicals were of biochemical
grade and purchased either from Sigma-Aldrich (St. Louis, MO) or from
Fisher Scientific (Pittsburgh, PA). The sulfated small molecules used
in this study were prepared earlier, as described in a series of articles,22 (link),24 (link)−33 (link) and were more than 95% pure. In these studies, purity was assessed
by a combination of techniques including HPLC/UPLC, HR-MS, and/or
elemental analysis.
fluorescein-EGR-fXIa (fEGR-fXIa)) was purchased from
Haematologic Technologies (Essex Junction, VT). Recombinant fXIa containing
only the catalytic domain (fXIa-CD) was a gift from Dr. Alireza Rezaie
(St. Louis University, MO). Chromogenic substrate S-2366 (
solutions of fXIa were prepared in 0.05 M Tris-HCl buffer, pH 7.4,
containing 0.15 M NaCl and 0.1% PEG8000. The buffer used in inhibition
studies was 0.05 M Tris-HCl buffer, pH 7.4, containing 0.15 M NaCl,
0.1% PEG8000, and 0.02% Tween80, while that used for all other studies
was devoid of 0.02% Tween80. All of the other chemicals were of biochemical
grade and purchased either from Sigma-Aldrich (St. Louis, MO) or from
Fisher Scientific (Pittsburgh, PA). The sulfated small molecules used
in this study were prepared earlier, as described in a series of articles,22 (link),24 (link)−33 (link) and were more than 95% pure. In these studies, purity was assessed
by a combination of techniques including HPLC/UPLC, HR-MS, and/or
elemental analysis.
10
Quantification of Activated Protein C
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PC determination was performed according to manufacturer’s instructions (Biophen Protein C, Aniara Diagnostica, OH) with minor modifications. Briefly, 25 μl of human control normal plasma (Biophen normal control plasma ref A223201) diluted 1:1 with sterile saline solution (0.9% NaCl) were incubated with 50 μl of protein C activator (Protac, 2 μM FP31W215A or 2 μM FP31WE) for 5 min at 37 °C in a 96-well plate. The amount of activated protein C was quantified by adding 125 μL of a solution of chromogenic substrate S-2366 (2.4 mM) (Diapharma Group, OH) and thrombin specific inhibitor argatroban (160 μM) (Sigma-Aldrich, MO) for 5 min at 37 °C. The reaction was quenched by addition of 150 μl citric acid and read at 405 nm. Mixing human control normal plasma with sterile saline solution to give 0, 25, 50 and 100% protein C generated standard calibration curves reported in Fig. 5A . Healthy control plasma was purchased from Innovative Research, MI and human abnormal control plasma depleted of protein C (ref A223301) was purchased from Biophen, Aniara Diagnostica, OH.
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