Anti-phosphorylated and pan, AKT, and MAPK antibodies, and PI3Kα/β/γ antibodies were purchased from Cell Signalling Technology (Cambridge, MA, USA). Anti-PI3Kδ antibody was purchased from R&D systems (Oxford, UK). Anti-CD138-PE, anti-CD90-FITC, anti-CD73-PE, anti-CD105-APC antibodies (Cat. 130-098-122, 130095403, 130095182, 130094926) and interleukin-6 (IL-6) were purchased from Miltenyi Biotec (Auburn, CA, USA). Idelalisib, CZC24832, duvelisib were obtained from Selleck Chemicals (Houston, TX, USA). All other reagents were obtained from Sigma-Aldrich (St Louis, MO, USA), unless otherwise indicated.
Interleukin 6 il 6
Manufactured by Miltenyi Biotec
Sourced in United States, Germany
Interleukin-6 (IL-6) is a cytokine that plays a crucial role in the regulation of the immune system and inflammatory response. It is a multifunctional protein involved in the activation and differentiation of various cell types, including T cells, B cells, and hematopoietic cells.
Automatically generated - may contain errors
Lab products found in correlation
Flt3 ligand flt3l, by Miltenyi Biotec (2 mentions)
Interleukin 3 il3, by Miltenyi Biotec (2 mentions)
Human serum, by Lonza (2 mentions)
Foetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Penicillin, by Thermo Fisher Scientific (2 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
Celltitre glo, by Promega (2 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions)
Idelalisib, by Selleck Chemicals (1 mentions)
Duvelisib, by Selleck Chemicals (1 mentions)
Anti cd73 pe, by Miltenyi Biotec (1 mentions)
Anti phosphorylated and pan, by Cell Signaling Technology (1 mentions)
Anti pi3kδ antibody, by R&D Systems (1 mentions)
Anti cd138 pe, by Miltenyi Biotec (1 mentions)
Anti cd90 fitc, by Miltenyi Biotec (1 mentions)
Czc24832, by Selleck Chemicals (1 mentions)
Karyomax colcemid solution, by Thermo Fisher Scientific (1 mentions)
Leishman stain, by Merck Group (1 mentions)
Trypsin edta 1, by Thermo Fisher Scientific (1 mentions)
Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (1 mentions)
7 protocols using interleukin 6 il 6
1
Multiparametric Analysis of PI3K Signaling
2
Bone Marrow Culture and Karyotyping Protocol
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The media and reagents used to culture BM were Dulbecco’s modified Eagle’s medium (DMEM; Invitrogen Corporation, Carlsbad, CA, USA), fetal bovine serum (FBS; JRS Scientific Inc., Woodland, CA, USA), penicillin/streptomycin (PAA Laboratories GmbH, Pasching, Australia), stem cell factor (SCF), interleukin-3 (IL-3), and interleukin-6 (IL-6) (Miltenyi Biotec, Bergisch Gladbach, Germany). The reagent, 1,4-BQ (CAS No: 106-51-4 and ≥ 98% purity) was obtained from Sigma-Aldrich Co. (St. Louis, MO, USA). Methylcellulose culture media were purchased from StemCell Technologies (Vancouver, BC, Canada) for culturing each progenitor lineages. The reagents used throughout the conventional karyotyping experiments were KaryoMAX Colcemid Solution, Trypsin EDTA 1× (Gibco, Grand Island, NY, USA), and Leishman stain (Sigma-Aldrich Co., St. Louis, MO, USA).
3
Umbilical Cord Blood CD34+ Cell Purification and Electroporation
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Umbilical CB samples were collected after normal deliveries, according to the institutional guidelines for discarded material. CB CD34+ cells were purified as previously described.3 (link) After immunomagnetic separation, CD34+ cells were seeded in 24-well plates at 5 × 105/ml in Iscove's modified Dulbecco's medium (IMDM) (GIBCO, Grand Island, NY, USA) containing 20% human serum (Bio-Whittaker, Walkersville, MD, USA), stem cell factor (SCF) (50 ng/ml), Flt3-ligand (FLT3L) (50 ng/ml), thrombopoietin (THPO) (20 ng/ml), interleukin-6 (IL6) (10 ng/ml) and interleukin-3 (IL3) (10 ng/ml) (all from Miltenyi Biotec, Auburn, CA, USA) and electroporated 24 hours later.
4
Biliary Differentiation of Stem Cells
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The siHBs were harvested using StemPro™ Accutase™ (Gibco, Waltham, MA, USA) and seeded onto collagen I-coated plates in Williams’E (Invitrogen, Waltham, MA, USA), 10% FBS, 1% BSA. The medium was then replaced by a biliary differentiation medium (BDM) with 25 ng/mL Growth Hormone (GH, Peprotech, Cranbury, NJ, USA) and 25 ng/mL EGF, for two days. The medium was replaced daily by BDM with 5 ng/mL Interleukin-6 (IL6, Miltenyi Biotec, Bergisch Gladbach, Germany) for seven days and until the end of the differentiation with 10 ng/mL EGF. For 3D differentiation, siChols were harvested using Accutase, and 2 × 105 cells were suspended in 100 µL 50:50 Matrigel (BD Biosciences, San Jose, CA, USA) and BDM on cell culture inserts with 1 µm pore size.
5
Cell viability assay in multiple myeloma
6
Evaluating Myeloma Cell Viability
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Antiphosphorylated and pan Akt antibodies
were purchased from Cell Signalling Technology (Cambridge, MA, USA).
Interleukin-6 (Il-6) was purchased from Miltenyi Biotec (Auburn, CA,
USA).
The authenticated multiple myeloma-derived cell line MM1
was obtained from the European Collection of Cell Cultures and was
cultured in RPMI 1640 medium supplemented with 10% foetal bovine serum,
penicillin, and streptomycin (all obtained from Invitrogen, Paisley,
UK).44 (link)Cell viability was determined
after 24 h using Cell Titre GLO (Promega,
Southampton, UK).31 (link) Data were normalized
to vehicle controls. All data points are represented as the mean with
s.e.
were purchased from Cell Signalling Technology (Cambridge, MA, USA).
Interleukin-6 (Il-6) was purchased from Miltenyi Biotec (Auburn, CA,
USA).
The authenticated multiple myeloma-derived cell line MM1
was obtained from the European Collection of Cell Cultures and was
cultured in RPMI 1640 medium supplemented with 10% foetal bovine serum,
penicillin, and streptomycin (all obtained from Invitrogen, Paisley,
UK).44 (link)Cell viability was determined
after 24 h using Cell Titre GLO (Promega,
Southampton, UK).31 (link) Data were normalized
to vehicle controls. All data points are represented as the mean with
s.e.
7
Isolation and Culture of CD34+ CB Cells
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Umbilical cord blood (CB) samples were collected after normal deliveries, according to the institutional guidelines for discarded material (Clearance of Ethical Commitee for Human experimentation of Modena: Secretary office Saverio Santachiara, santachiara.saverio@policlinico.mo.it, approval date: 18.01.2005; approval file number # 793/CE). CB CD34+ cells were purified as previously described (13) . After immunomagnetic separation, CD34+ cells were seeded in 24-well plates at 5 × 10 5 /ml in Iscove's modified Dulbecco's medium (IMDM) (Euroclone SPA, Milan, Italy) containing 20% human serum (Bio-Whittaker, Walkersville, MD, USA), stem cell factor (SCF) (50 ng/ml), FLT3-ligand (FLT3L) (50 ng/ml), thrombopoietin (TPO) (20 ng/ml), interleukin-6 (IL6) (10 ng/ml), and interleukin-3 (IL3) (10 ng/ml) (all from Miltenyi Biotec, Auburn, CA, USA).
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