A pooled single cell suspension of splenocytes and lymph node cells prepared as described above were re-suspended in Iscove’s Modified Dulbecco’s Media (Sigma-Aldrich) supplemented with 10% heat inactivated fetal bovine serum, 4 mM glutamine, 50 µM β-mercaptoethanol, 100 U/ml penicillin, and 0.1 mg/ml streptomycin (Sigma-Aldrich). 2 × 105 cells of leukocytes were cultured for 72 h and stimulated with 50 µg/ml mBSA or 1 µg/ml anti-CD3 antibodies in the presence or absence of 500 U/ml of IFN-α. After 72 h, cells were harvested and analyzed for CD4, CD25, and Foxp3 expression as described above.
Iscove s modified dulbecco s media
Manufactured by Merck Group
Sourced in United States
Iscove's Modified Dulbecco's Media is a cell culture medium formulation designed to support the growth and maintenance of a wide range of cell types. It is a modification of the original Dulbecco's Modified Eagle's Medium, providing additional nutrients and supplements to enhance cell viability and proliferation.
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Lab products found in correlation
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Amphotericin b, by Thermo Fisher Scientific (2 mentions)
L glutamine, by Merck Group (2 mentions)
Mda mb 231, by Merck Group (2 mentions)
Streptomycin, by Merck Group (1 mentions)
Penicillin, by Merck Group (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Glutamax, by Thermo Fisher Scientific (1 mentions)
Fetal calf serum (fcs), by Thermo Fisher Scientific (1 mentions)
Mycoalert, by Lonza (1 mentions)
Wild type c57bl 6j mice, by Jackson ImmunoResearch (1 mentions)
Gentamicin, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Corning (1 mentions)
Penicillin, by Corning (1 mentions)
Penicillin streptomycin, by Merck Group (1 mentions)
Amphotericin b, by Merck Group (1 mentions)
Leibovitz l 15 media, by Merck Group (1 mentions)
Mcf 10a, by American Type Culture Collection (1 mentions)
5 protocols using iscove s modified dulbecco s media
1
Isolation and Stimulation of Murine Lymphocytes
2
HEK293T Cell Culture Conditions
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HEK293T cells (Human embryonic kidney, Lehner laboratory stocks) were cultured in Iscove’s Modified Dulbecco’s Media (Sigma) supplemented with 1% GlutaMAX, 10% FCS, 100 units/ml penicillin, and 100 µg/ml streptomycin (all from Thermo Fisher), at 37 °C in 5% CO2. Cells were confirmed mycoplasma negative (MycoAlert, Lonza).
3
C57BL/6J Mice S. stercoralis Infection
4
Culturing Cancer and MSC Cell Lines
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Cancer cell lines MDA-MB-231, MCF-7 and SW837 were obtained from the Russian cell culture collection (Russian Branch of the ETCS, Russia, St. Petersburg). MSC cells were kindly gifted by Dr. Matveeva [3] (link). MCF-7 and SW837 cells were cultivated in Iscove's modified Dulbecco's media (Sigma) with 10% FBS (Gibco BRL Co., Gaithersburg, MD), 2 mM L-glutamine (Sigma), 250 mg/mL amphotericin B and 100 U/mL penicillin/streptomycin (GIBCO BRL Co., Gaithersburg, MD). MDA-MB-231 cells were cultivated in Leibovitz (L15) media (Sigma) supplemented with 10% FBS, 2 mM L-glutamine, 250 mg/ml amphotericin B and 100 U/mL penicillin/streptomycin. Cells were grown in a humidified atmosphere of 5% CO2 in air at 37°C and were passaged with 0.05% trypsin-EDTA every 3–4 days.
5
Cell Culture Protocols for Cancer and Normal Cells
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African green monkey kidney fibroblasts (CV-1) and non-tumorigenic human breast epithelial cells (MCF 10A) were obtained from the American Type Culture Collection (ATCC; Manassas, VA). RLS cells were generously provided by Dr. V. I. Kaledin (Institute of Cytology and Genetics SB RAS). Cancer cell lines MDA-MB-231, MCF-7, A549, U87MG, BT549 and BT20 were obtained from the Russian cell culture collection (Russian Branch of the ETCS, St. Petersburg, Russia). MDA-MB-231 cells were grown in Leibovitz media (L15, Sigma-Aldrich) supplemented with 10% fetal bovine serum (FBS, HyClone, USA), 2mM L-glutamine, 250 mg/ml amphotericin B and 100 U/ml penicillin/streptomycin. Other cancer cells were cultivated in Iscove's modified Dulbecco's media (Sigma) with 10% FBS (Gibco BRL Co., Gaithersburg, USA), 2mM L-glutamine (Sigma-Aldrich), 250 mg/mL amphotericin B and 100 U/ml penicillin/streptomycin (GIBCO BRL Co., Gaithersburg, USA). MCF 10A were cultured in MEGM BulletKit (Lonza/Clonetics Corporation, USA). CV-1 cells were grown in Dulbecco's modified Eagle's mediun (DMEM, Invirtogen, USA) supplemented with 10% of FBS and antibiotics (100 U/ml penicillin/streptomycin).
Cells were grown in a humidified atmosphere of 5% CO2 in air at 37°C and were passaged with 0.05% trypsin-EDTA every 2–4 days.
Cells were grown in a humidified atmosphere of 5% CO2 in air at 37°C and were passaged with 0.05% trypsin-EDTA every 2–4 days.
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