Arginine deprivation was carried out as described previously by Pawar et al. (12 (link)). Briefly, mid-log-phase promastigotes (1 × 107 cells/ml) were washed with Earle’s balanced salt solution twice and resuspended in arginine-deficient medium M-199 (Biological Industries Ltd.) at 26°C for the specified period before being transferred to ice. Arginine-deprived cells were washed twice with ice-cold Earle’s balanced salt solution before being used for transport assays and Northern and Western blot analyses.
M 199
Manufactured by Sartorius
Sourced in Israel
The M-199 is a high-precision electronic microscale from Sartorius designed for laboratory applications. It offers accurate weighing capabilities with a capacity of up to 220 grams and a readability of 0.1 milligrams. The scale features a compact, space-saving design and an intuitive user interface.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Sartorius (4 mentions)
Vitamin b12, by Merck Group (3 mentions)
Roswell park memorial institute (rpmi), by Sartorius (3 mentions)
Chir99021, by Bio-Techne (3 mentions)
Glutamine, by Sartorius (2 mentions)
B27 insulin, by Thermo Fisher Scientific (2 mentions)
Roswell park memorial institute (rpmi), by Bio-Techne (2 mentions)
B27 insulin, by Bio-Techne (2 mentions)
L glutamine, by Sartorius (2 mentions)
L glutamine, by Bio-Techne (2 mentions)
Penicillin streptomycin solution, by Sartorius (1 mentions)
Earle s salts, by Sartorius (1 mentions)
Retinoic acid, by Merck Group (1 mentions)
Accutase, by STEMCELL (1 mentions)
Nutristem, by Sartorius (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Basic fibroblast growth factor (bfgf), by Thermo Fisher Scientific (1 mentions)
6 protocols using m 199
1
Axenic Differentiation of L. donovani Promastigotes
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L. donovani MHOM/SD/00/1S (24 (link)) promastigotes were grown in medium M-199, Earle’s salts (Biological Industries Ltd.) supplemented with 10% heat-inactivated fetal bovine serum (FBS) (Gibco Ltd.) and a 1% penicillin-streptomycin solution (Biological Industries Ltd.). Axenic differentiation of L. donovani promastigotes to amastigotes was carried out as described previously (7 (link)). Briefly, mid-log-phase promastigotes were washed twice in Earle’s salt solution and finally suspended in amastigote medium containing M-199 at pH 5.5 supplemented with 0.5 mM sodium succinate, 25% bovine serum, and 1% penicillin-streptomycin solution. Mature amastigotes developed at 5 days after exposure of promastigotes to the differentiation medium.
Arginine deprivation was carried out as described previously by Pawar et al. (12 (link)). Briefly, mid-log-phase promastigotes (1 × 107 cells/ml) were washed with Earle’s balanced salt solution twice and resuspended in arginine-deficient medium M-199 (Biological Industries Ltd.) at 26°C for the specified period before being transferred to ice. Arginine-deprived cells were washed twice with ice-cold Earle’s balanced salt solution before being used for transport assays and Northern and Western blot analyses.
+ Expand
Arginine deprivation was carried out as described previously by Pawar et al. (12 (link)). Briefly, mid-log-phase promastigotes (1 × 107 cells/ml) were washed with Earle’s balanced salt solution twice and resuspended in arginine-deficient medium M-199 (Biological Industries Ltd.) at 26°C for the specified period before being transferred to ice. Arginine-deprived cells were washed twice with ice-cold Earle’s balanced salt solution before being used for transport assays and Northern and Western blot analyses.
2
Comparative Evaluation of Viral RNA Extraction
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Archived nasopharyngeal samples (including Bronchoalveolar lavage (BAL) (N = 8), tracheal aspirations (N = 32) or nasopharyngeal swabs (N = 403)) of patients hospitalized at Chaim Sheba Medical Center, collected into Virocult liquid viral transport medium (LVTM) (Medical Wire & Equipment Co, Wiltshire, United Kingdom) and stored at -70°C, were used to evaluate the performance of the three extraction systems. Representative samples covering the range of cycle threshold values (Ct) below 35, which were previously extracted by easyMAG and tested for the common human respiratory viruses, were selected.
The archived samples were thawed and the volume of the sample was adjusted with M199 (transport medium Biological Industries (BI), Beit Haemek, Israel), compatible with all three systems. The archived samples (N = 262) included positive samples for influenza A (N = 47), H1N1pdm (N = 22), influenza B (N = 21), RSV (N = 21), hMPV (N = 23), parainfluenza-3 (N = 20) and adenovirus (N = 23), as well as negative control samples (N = 181).
The technical and physical properties of the three platforms, including platform size, nucleic acid collection options (tubes/plate/cartridge), sample loading method (directly/indirectly from the sample tube), the existence of a barcode reader and user operation convenience, are summarized inTable 1 .
The archived samples were thawed and the volume of the sample was adjusted with M199 (transport medium Biological Industries (BI), Beit Haemek, Israel), compatible with all three systems. The archived samples (N = 262) included positive samples for influenza A (N = 47), H1N1pdm (N = 22), influenza B (N = 21), RSV (N = 21), hMPV (N = 23), parainfluenza-3 (N = 20) and adenovirus (N = 23), as well as negative control samples (N = 181).
The technical and physical properties of the three platforms, including platform size, nucleic acid collection options (tubes/plate/cartridge), sample loading method (directly/indirectly from the sample tube), the existence of a barcode reader and user operation convenience, are summarized in
3
Cardiac Differentiation of iPSCs
4
Cardiomyocyte Differentiation Protocol
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Prior to differentiation, cells were dissociated with Accutase™ (StemCell Technologies, Vancouver, BC, Canada) and passaged to six-well plates. Until 100% confluence of iPSCs, NutriStem™ (Biological Industries) was refreshed. On day 0, the medium was changed to 3 mL of RPMI (Biological Industries), supplemented with 0.5% L-glutamine (Biological Industries), B27-Insulin (Invitrogen, Carlsbad, CA, USA) and 4.5 µM CHIR-99021 (Tocris, Bristol, UK). On day 2, the medium was changed to 3 mL of RPMI supplemented with 0.5% L-glutamine, B27-Insulin, and 5 µM IWP-2 (Tocris). On day 4, the medium was changed to 3 mL of RPMI supplemented with 0.5% L-glutamine and B27-Insulin. This medium was refreshed on day 6. On day 8, the medium was changed to 3 mL of RPMI supplemented with 0.5% L-glutamine and B27, and this medium was refreshed on day 10. From day 12, the medium was changed to M-199 (Biological Industries), supplemented with 0.1% penicillin/streptomycin, 5% fetal bovine serum (FBS, Biological Industries), 0.6 mM CuSO4·5 H2O, 0.5 mM ZnSO4·7 H2O, and 1.5 mM Vitamin B12 (Sigma-Aldrich, Darmstadt, Germany). This medium was refreshed every other day.
5
Culturing Human Umbilical Vein Endothelial Cells
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Human umbilical vein endothelial cells (HUVECs) (kindly provided by Prof. Gera Neufeld, Technion, Israel) were grown on 10 cm plates, coated with 0.2% gelatin in Dulbecco’s phosphate-buffered saline (PBS; Biological Industries, Israel) and overlaid with growth medium comprised of Earle’s salt base (M-199) medium supplemented with 20% fetal bovine serum (FBS), 1% antibiotics, 1% vitamins, and glutamine (Biological Industries, USA) and freshly added basic fibroblast growth factor (bFGF) (PeproTech, Israel) (5 ng/ml). Jurkat T cells (kindly provided by Prof. Debbie Yablonski, Technion, Israel) were maintained in RPMI-1640 (Gibco–Life Technology, USA) with high glucose, 10% heat inactivated FBS, and 1% antibiotics. All cells were incubated at 37°C, 5% CO2 incubator.
6
Cardiac Differentiation of Induced Pluripotent Stem Cells
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Prior to differentiation,
cells were passed to 6-well plates. NutriStem was refreshed daily
until iPSCs reached 100% confluence. At that point (Day 0), the medium
was changed to 3 mL of RPMI (Biological Industries), supplemented
with 0.5%l -glutamine (Biological Industries), B27-Insulin
(Invitrogen, Carlsbad, California), and 4.5 μM CHIR-99021 (Tocris,
Bristol, UK). On Day 2, the medium was changed to 3 mL of RPMI supplemented
with 0.5%l -glutamine, B27-Insulin, and 5 μM IWP-2
(Tocris). On Day 4, the medium was changed to 3 mL of RPMI supplemented
with 0.5%l -glutamine and B27-Insulin, and this medium was
refreshed on Day 6. On Days 8 and 10, the medium was changed to 3
mL of RPMI supplemented with 0.5%l -glutamine and B27. From
Day 12, the medium was changed to M-199 (Biological Industries), supplemented
with 0.1% penicillin/streptomycin, 5% fetal bovine serum (FBS, Biological
Industries), 0.6 mM CuSO4·5H2O, 0.5 mM
ZnSO4·7H2O, and 1.5 mM Vitamin B12 (Sigma-Aldrich).
This medium was refreshed every other day.74 (link)
cells were passed to 6-well plates. NutriStem was refreshed daily
until iPSCs reached 100% confluence. At that point (Day 0), the medium
was changed to 3 mL of RPMI (Biological Industries), supplemented
with 0.5%
(Invitrogen, Carlsbad, California), and 4.5 μM CHIR-99021 (Tocris,
Bristol, UK). On Day 2, the medium was changed to 3 mL of RPMI supplemented
with 0.5%
(Tocris). On Day 4, the medium was changed to 3 mL of RPMI supplemented
with 0.5%
refreshed on Day 6. On Days 8 and 10, the medium was changed to 3
mL of RPMI supplemented with 0.5%
Day 12, the medium was changed to M-199 (Biological Industries), supplemented
with 0.1% penicillin/streptomycin, 5% fetal bovine serum (FBS, Biological
Industries), 0.6 mM CuSO4·5H2O, 0.5 mM
ZnSO4·7H2O, and 1.5 mM Vitamin B12 (Sigma-Aldrich).
This medium was refreshed every other day.74 (link)
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