LNCaP-abl cells were resuspended, dissociated into single cells and were cultured in 6-well ultra-low attachment culture plates (Corning, Amsterdam, The Netherlands) with DMEM/F-12 (Gibco, Carlsbad, CA) supplemented with 1% L-Glutamax, 1% P/S, 20 ng/μL human epidermal growth factor (hEGF; Gibco), 20 ng/μL basic fibroblasts growth factor (bFGF; Gibco), 1× B27 without vitamin A (Invitrogen), heparin sodium, 1× insulin-transferrin-selenium A (Invitrogen), 1× non-essential amino acids (Invitrogen), 0.1 mM β-mercaptoethanol (Sigma) and 103 U/mL leukemia inhibitory factor (LIF; Millipore, Billerica, MA). Besides these, the concentration of E2 was 1 nM in the control group and 10 nM in the E2 group. Sphere cultures were seeded at a density of 2 × 103 cells/mL and culture media was fully replaced every 96 h. Parental and prostasphere cultures were propagated for 12 days before the spheres were enzymatically dissociated using StemPro Accutase (Gibco) for second and third generation cell culture for 7 days, separately. Spheres were imaged and visualized via Olympus CX41 microscopy to evaluate the volume, or were collected for qRT-PCR or immunofluorescence detection.
Sodium heparin
Manufactured by Thermo Fisher Scientific
Sourced in United States, Canada, Belgium
Sodium heparin is an anticoagulant used in laboratory settings to prevent blood from clotting during sample collection and analysis. It functions by inhibiting the activation of the coagulation cascade, a series of enzymatic reactions that lead to the formation of a fibrin clot.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (11 mentions)
Penicillin, by Thermo Fisher Scientific (7 mentions)
Streptomycin, by Thermo Fisher Scientific (7 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (4 mentions)
4 6 diamidino 2 phenylindole (dapi), by Beyotime (3 mentions)
Histopaque 1083, by Merck Group (3 mentions)
Sulforhodamine b srb, by Thermo Fisher Scientific (2 mentions)
Distearoyl sn glycero 3 phosphoethanolamine peg dspe peg, by Merck Group (2 mentions)
Collagenase type 4, by Thermo Fisher Scientific (2 mentions)
Roswell park memorial institute (rpmi), by Thermo Fisher Scientific (2 mentions)
Ack lysis buffer, by Lonza (2 mentions)
Ack lysing buffer, by Thermo Fisher Scientific (2 mentions)
Dianisidine dihydrochloride, by Merck Group (2 mentions)
Glucose oxidase peroxidase enzyme, by Merck Group (2 mentions)
3 hydroxybutyrate dehydrogenase, by Roche (2 mentions)
B27 without vitamin a, by Thermo Fisher Scientific (1 mentions)
Insulin transferrin selenium a, by Thermo Fisher Scientific (1 mentions)
Cx41 microscopy, by Olympus (1 mentions)
Leukemia inhibitory factor, by Merck Group (1 mentions)
Non essential amino acids (neaa), by Thermo Fisher Scientific (1 mentions)
26 protocols using sodium heparin
1
Culturing Prostaspheres from LNCaP-abl Cells
2
CNT-DOX Nanocarrier for Cancer Therapy
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Single-walled CNTs (purity 0.90%) were purchased from Shenzhen Nanotech Port (Shenzhen, People’s Republic of China). DOX (production date: 2014/05/11, purity >98%) was obtained from Beijing Yi-He Biotech Co. Ltd. 1-MA, distearoyl-sn-glycero-3-phosphoethanolamine-PEG (DSPE-PEG), and dimethyl sulfoxide were obtained from Sigma-Aldrich Co. LLC. Sulforhodamine B, Dulbecco’s Modified Eagle’s Medium (DMEM) cell culture medium, penicillin, streptomycin, fetal bovine serum (FBS), and heparin sodium were bought from Gibco Invitrogen. DAPI and hematoxylin and eosin (H&E) were supplied by Beyotime Biotechnology Co. Ltd. Other reagents were acquired from China National Medicine Corporation Ltd. Also, 808 nm laser (diode laser, 2 W/cm2, CW) was supplied by Changchun Laser Research Center. The dialysis bags (MWCO =10,000) were obtained from Spectrum Laboratories Inc.
3
MoS2-based Doxorubicin Delivery System
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Molybdenum disulfide (MoS2, purity > 95%) were purchased from DK nano Co. Ltd Beijing. Doxorubicin (DOX, 20120503, purity > 98%) was gotten from Beijing Yi-He Biotech Co. Ltd. Thioglycollic acid (HSCH2COOH, 70% in water), NH2-PEG2000-OCH3, N-(3-dimethylamino propyl-N0-ethylcar-bodiimide) hydrochloride (EDC∙HCl), IR 783 dye and dimethyl sulfoxide (DMSO) were obtained from Sigma-Aldrich Co. LLC. Sulforhodamine B (SRB), RPMI 1640 cell culture medium, penicillin, streptomycin, fetal bovine serum (FBS), and heparin sodium were bought from Gibco Invitrogen.
4
Doxorubicin-Loaded Lipid Nanoparticles
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Doxorubicin (DOX, 20,170,511, purity >98%) was gotten from Beijing Yi-He Biotech Co. Ltd. All single-stranded DNAs were purchased from Takara Bio Co. (Dalian, China), and the specific sequences were displayed in Table 1 . Phospholipids, 1,2-dioleoy-sn-glycero-3-phosphoethanolamine (DOPE), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC) and cholesteryl hemisuccinate (CHEMS) were purchased from Avanti Polar Lipids (Alabama, USA), and N-(carbonyl-methoxy-polyethylene-glycol-2000)-1,2-distearoyl-sn-glycero-3-phosphoethanolame (DSPE-mPEG2000) from Lipoid (Steinhausen, Switzerland). Cholesterol and calcein were obtained from Sigma-Aldrich Ltd (Auckland, New Zealand). PCR primer, loading Buffer, and Golden View was obtained from Beijing Ding Guo Chang Sheng Biotechnology Co. Ltd. Gel Extraction Kit, 2× Taq Master Mix, and 50× TAE was obtained from Beijing Com Win Biotech Co. Ltd. Sulforhodamine B (SRB), DMEM cell culture medium, penicillin, streptomycin, fetal bovine serum (FBS), and heparin sodium were bought from Gibco Invitrogen. DAPI, hematoxylin and eosin were supplied by Beyotime Biotechnology Co. Ltd. Other reagents were acquired from China National Medicine Corporation Ltd.
5
Biofilm Formation of E. faecalis
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Overnight cultures in GM17 of E. faecalis were adjusted to OD600 of 0.2 in fresh M17 supplemented with 2% glucose. One hundred microliters of the bacterial suspension were inoculated into CytoOne polystyrene microwells plate coated with 1 μg/ml of hyaluronic acid (Sigma-Aldrich), chondroitin sulfate (Carl Roth, Karlsruhe, Germany), or heparin sodium (ThermoFisher). After 24 or 48 h of incubation at 37°C, the plates were washed with 0.9% NaCl to remove unbound bacteria. Each well was then stained with 0.1% (wt/vol) crystal violet for 15 min at room temperature. Wells were then rinsed two times with 0.9% NaCl. Adherent cells were dissolved in 30% acetic acid, and the OD550 was measured using a microplate reader Nano Tecan (Life Science). At least three experiments were performed for each condition.
6
Synthesis and Characterization of Gadolinium-Ferritin Nanoparticles
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DOX (purity >98%) was obtained from Beijing Yi-He Biotech Co., Ltd., Beijing, People’s Republic of China. GdCl3 (purity >99.9, wt.%) was purchased from Sinopharm Chemical Reagent Co., Ltd. (Shanghai, People’s Republic of China). Ferritin from equine spleen, FA, citric acid, polyethylenimine (PEI; molecular weight =2,500), diethylene glycol, ethylenediamine, formamide, N-hydroxysuccinimide (NHS), N-(3-dimethylaminopropyl-N′-ethylcarbodiimide) hydrochloride (EDC-HCl), and dimethyl sulfoxide were obtained from Sigma-Aldrich Co. (St Louis, MO, USA). 3-(4,5-Dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide (MTT), Dulbecco’s Modified Eagle’s Medium cell culture medium, penicillin, streptomycin, fetal bovine serum, and heparin sodium were purchased from Thermo Fisher Scientific (Waltham, MA, USA). Other reagents were acquired from China National Medicines Corporation Ltd. The dialysis bags (molecular weight cutoff [MWCO] =10,000) were obtained from Spectrum Laboratories, Inc. Ultrapure water was used throughout the experiments.
7
Photodynamic Therapy Drug Formulations
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DOX (20140511, purity >98%) and HMME (20160302, purity >98%) were obtained from Beijing Yi-He Biotech Co. Ltd (Beijing, People’s Republic of China). Dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), cholesterol, distearoyl-sn-glycero-3-phosphoethanolamine-PEG (DSPE-PEG), and dimethyl sulfoxide were obtained from Sigma-Aldrich Co. (St Louis, MO, USA). Sulforhodamine B (SRB), DMEM cell culture medium, penicillin, streptomycin, fetal bovine serum (FBS), and heparin sodium were bought from Thermo Fisher Scientific (Waltham, MA, USA). 4′,6-diamidino-2-phenylindole (DAPI) and hematoxylin and eosin were supplied by Beyotime Biotechnology Co. Ltd (Shanghai, People’s Republic of China). Other reagents were acquired from China National Medicine Corporation Ltd (Chongqing, People’s Republic of China). A 532 nm laser (diode laser, 0–300 mW/cm2, CW) was supplied by Changchun Laser Research Center (Changchun, People’s Republic of China).
8
Isolation of Immune Cells from Tissues
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Cervical LN or spleens were dissociated with the end of a 3 mL syringe plunger through 70 μM nylon mesh in RPMI (Life Technologies, Waltham, MA) supplemented with 10% FBS, 100 U/mL penicillin, 100 μg/mL streptomycin, and 50 μM β-mercaptoethanol (complete RPMI). Cells were treated with ACK lysis buffer (Lonza, Mapleton, IL) to remove red blood cells (RBC). To isolate cells from the lacrimal or salivary glands, glands were incubated in Collagenase type IV (Life Technologies, Waltham, MA) at 37°C with shaking for 1 or 1.5 hours, respectively. Glands were then dissociated with the end of a 3 mL syringe plunger through 40 μM nylon mesh and RBC lysed as above to obtain single cell suspensions. Peripheral blood samples were collected through the retro-orbital plexus and diluted in PBS containing 20 U/mL heparin sodium (Fisher Scientific, Hampton, NH). Single cell suspensions were obtained using Histopaque-1083 (Sigma-Aldrich, St. Louis, MO) according to the manufacturer’s instructions.
9
Heparin Infusion Mitigates Insulin Site-Loss
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Example 7
The objectives of this study include examining site-loss mitigation when heparin is used in continuous infusion along with insulin. U100 of an insulin Humalog was added with 4 mg/mL heparin sodium (purchased from Fisher™, 193 U/mg) and filtered. The actual heparin concentration was 3.55 mg/mL or 685 U/mL. The dosing scheme for pigs 3 and 4 (IM3 and IM4, respectively) are shown in Table 11 below. Based on the glucose monitoring results for IM3 and IM4 (shown in
10
Isolation of Immune Cells from Tissues
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