We obtained calcium chloride (CaCl2), sodium chloride (NaCl), and alginic acid sodium salt powder from Sigma-Aldrich (Tokyo, Japan). Sucrose was purchased from China National Pharmaceutical Group Co., Ltd. (Sinopharm, Beijing, China). Fibrinogen and thrombin were purchased from Beijing Solarbio Science and Technology Co., Ltd. (Beijing, China). All chemicals were used without further purification. Three types of non-steroidal anti-inflammatory drugs (aspirin, indomethacin, and nimesulide) were purchased from Sigma-Aldrich (Tokyo, Japan).
Fibrinogen
Manufactured by Solarbio
Sourced in China
Fibrinogen is a laboratory-grade reagent used in various research and clinical applications. It is a soluble plasma glycoprotein that plays a crucial role in the blood clotting process. Fibrinogen is a essential component in the formation of fibrin, which is the main structural protein in blood clots.
Automatically generated - may contain errors
Lab products found in correlation
Thrombin, by Solarbio (4 mentions)
Nimesulide, by Merck Group (1 mentions)
Aspirin, by Merck Group (1 mentions)
Sucrose, by Sinopharm (1 mentions)
Alginic acid sodium salt powder, by Merck Group (1 mentions)
Indomethacin, by Merck Group (1 mentions)
Agarose, by Sinopharm (1 mentions)
Precision plus protein dual color standard, by Bio-Rad (1 mentions)
Urokinase, by Maclin Biochemical Technology (1 mentions)
Triethylamine, by Solarbio (1 mentions)
Sodium acetate, by Sinopharm (1 mentions)
Hydrochloric acid (hcl), by Sinopharm (1 mentions)
Sodium hydroxide (naoh), by Sinopharm (1 mentions)
Sodium bicarbonate, by Sinopharm (1 mentions)
Perchloric acid, by Sinopharm (1 mentions)
Trichloroacetic acid, by Sinopharm (1 mentions)
Tris hcl, by Sinopharm (1 mentions)
Hplc grade methanol meoh, by Thermo Fisher Scientific (1 mentions)
Academic ultrapure water system, by Merck Group (1 mentions)
Fibrinogen, by Merck Group (1 mentions)
6 protocols using fibrinogen
1
Biomaterial Synthesis with Bioactive Compounds
2
Purification and Characterization of Urokinase
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Ammonium sulfate (NH4)2SO4), Tris (hydroxymethyl) aminomethane, agarose, and ethylene diamine tetraacetic acid (EDTA) were purchased from Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China). Urokinase was purchased from Shanghai Macklin Biochemical Technology Co., Ltd. (Shanghai, China). Fibrinogen was purchased from Beijing Solarbio Science & Technology Co., Ltd. (Beijing, China). Thrombin was purchased from Abmole (Houston, TX, USA). An Omni-EasyTM One-step PAGE Gel Fast Preparation Kit (12.5%, Catalog No. PG213) was purchased from Shanghai Epizyme Biomedical Technology Co., Ltd. (Shanghai, China). A 10× Tris/Glycine/SDS Buffer (Catalog 1610732), 4× Laemmli Sample Buffer (Catalog No. 1610747) and Precision Plus Protein Dual Color Standards with MW of 10–250 kDa (Catalog 1610374) were purchased from Bio-Rad Laboratories Inc. (Hercules, CA, USA). The bicinchoninic acid (BCA) protein quantitation kit (Catalog PA115-01) was purchased from Tiangen Biotech (Beijing, China) Co., Ltd. (Beijing, China). DEAE-Sepharose FF, Sephadex G-50, soybean trypsin inhibitor (SBTI), and phenyl methane sulfonyl fluoride (PMSF) were purchased from Shanghai yuanye Bio-Technology Co., Ltd. (Shanghai, China). Epsilon amino caproic acid (EACA) was purchased from Shanghai Aladdin Biochemical Technology Co., Ltd. (Shanghai, China). All reagents used were of analytical grade unless otherwise stated.
3
Quantification of Amino Acids and Biogenic Amines in Soybeans
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Soybeans were from Heilongjiang Province, China. The standards of 17 FAAs (His, Tyr, Arg, Val, Met, Phe, Ile, Leu, Ser, Gly, Ala, Thr, Lys, Asp, Glu, Cys, Pro) and eight BAs (putrescine, cadaverine, tyramine, spermidine, spermine, histamine, and 2-phenylethylamine), tetrahydrofuran, triethylamine, thrombin, and fibrinogen were bought from Solarbio (Beijing, China). Hydrochloric acid, sodium acetate, trichloroacetic acid, sodium bicarbonate, dansyl chloride, perchloric acid, sodium hydroxide, Tris-HCl, and ammonia were procured from Sinopharm Chemical Reagent Co., Ltd., Shanghai, China. All the above reagents were of analytical grade. 2-Methyl-3-heptanone was of chromatographic grade and bought from Solarbio (Beijing, China). Methanol and acetonitrile were of chromatographic grade and purchased from Beijing Bailingwei Technology Co., Ltd., Beijing, China.
4
3D Tumor Spheroid Formation Protocol
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The 5 μl thrombin (0.1 U/μl) (Sea Run Holdings, Maine, USA) were added into the 24-well plates. Then a blend of the cell suspension and an equal volume of the fibrinogen (Solarbio, Beijing, China) was seeded into the 24-well plate (1000 cells/well) and mixed with thrombin. The plates were placed in incubator for 10 min until the mixture into semi-solid and 1 ml culture medium was replenish. After incubation for 7 days, the tumor spheres were formed and analyzed.
5
Phytochemical Analysis of Earthworm Powder
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Hypoxanthine, uridine, adenine, inosine, guanosine, and adenosine were purchased from Chengdu Desite Bio-Technology Co., Ltd. (Chengdu, China). Phenylalanine and tryptophan were bought from Shanghai Aladdin Biochemical Technology Co., Ltd (Shanghai, China). Potassium dihydrogen phosphate (KH2PO4) was purchased from Tianjin Fengchuan Chemical Reagent Co., Ltd. Thrombin, fibrinogen, and phosphate buffer saline (PBS, pH 7.2–7.4, 0.01 M) were purchased from Beijing Solarbio Science and Technology Co., Ltd. (Beijing, China). HPLC-grade methanol (MeOH) was obtained from Fisher (Leicestershire, UK). Ultrapure water was obtained from a Milli-Q academic ultrapure water system (Millipore, Milford, MA, USA).
Forty batches of Pheretimasamples were purchased from medicine markets and pharmacies in different provinces in China. The Pheretima samples were dried at 60°C under reduced pressure before being pulverized into homogeneous powder (through a 65-mesh sieve).
Although the detailed information on Pheretima species was not identified because they have been processed into decoction pieces. The samples were authenticated as Pheretima according to Chinese Pharmacopeia by Prof. Yanxu Chang (Tianjin University of Traditional Chinese Medicine).
Forty batches of Pheretimasamples were purchased from medicine markets and pharmacies in different provinces in China. The Pheretima samples were dried at 60°C under reduced pressure before being pulverized into homogeneous powder (through a 65-mesh sieve).
Although the detailed information on Pheretima species was not identified because they have been processed into decoction pieces. The samples were authenticated as Pheretima according to Chinese Pharmacopeia by Prof. Yanxu Chang (Tianjin University of Traditional Chinese Medicine).
6
Generating Interstitial Flow in Fibrin Matrix
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Fibrinogen (F8630; Sigma-Aldrich, USA) was dissolved in 0.9% NaCl at a concentration of 10 mg/mL and stored at −20 °C until use. The stored Fibrinogen was thawed and diluted with Dulbecco’s phosphate-buffered saline (D-PBS; Solarbio; Beijing, China) from 10 mg/mL to 2.5 mg/mL, filter-sterilized (0.22 μm pore) and supplemented with aprotinin (0.45 TIU/mL, A1153; Sigma-Aldrich; USA). Then, the diluted Fibrinogen was mixed with thrombin (1 U/mL, T4648; Sigma-Aldrich; USA) and immediately introduced into the channels C1, C3, and C5 via inlet 1, 3, and 5, and left to gel for 10 min at room temperature. After fibrin gelling, the cell culture medium was introduced into channels C2 and C4 via inlets 2 and 4. Different volumes of medium were added into C2 and C4 to generate IF in the fibrin matrix across channel C3. IF flowing from channel C2 to channel C4 was termed upstream IF, while the IF in the reverse direction was termed downstream IF. The medium in inlets C2 and C4 was changed every 12 h to maintain the pressure gradient between C2 and C4.
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