White PCL particles with the molecular weight of 80,000 g/mol was supplied by Sigma (Shanghai, China). Phosphate Buffered Saline (PBS), acetone, and hexafluoro-isopropanol (HFIP) was purchased from China National Medicines Corporation Ltd., (Shanghai, China). SIM was purchased from Aladdin (Shanghai, China). nHA was purchased from McLean (Shanghai, China). BS350A CCK-8 solution was supplied from Labgic Technology Co., Ltd (Beijing, China), Primary human bone marrow derived mesenchymal stem cells (hBMSCs) and fetal bovine serum (FBS) were supplied from Sciencell (San Diego, CA, United States), alpha minimum essential medium (α-MEM) and antibiotics were obtained from Thermo Fisher Scientific (Rockford, IL, United States).
Alpha minimum essential medium α mem
Manufactured by Thermo Fisher Scientific
Sourced in United States, United Kingdom
Alpha‐minimum essential medium (α‐MEM) is a cell culture medium formulation used to support the growth and maintenance of various cell types in vitro. It contains a balanced salt solution, amino acids, vitamins, and other essential components required for cell proliferation and survival.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (12 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (3 mentions)
Fetal bovine serum (fbs), by Biowest (3 mentions)
Gst rankl, by Oriental Yeast (2 mentions)
Recombinant human m csf, by R&D Systems (2 mentions)
Tetrahydrofuran, by Thermo Fisher Scientific (2 mentions)
Human lactoferrin, by Merck Group (2 mentions)
Acetone, by Thermo Fisher Scientific (2 mentions)
Phosphate buffered saline tablet, by Merck Group (2 mentions)
Dimethyl sulfoxide (dmso), by Thermo Fisher Scientific (2 mentions)
Granulocyte macrophage colony stimulating factor gm csf, by Thermo Fisher Scientific (2 mentions)
Nile red dye nr, by Merck Group (2 mentions)
Antibiotic antimycotic solution 100x, by Merck Group (2 mentions)
Micro bca protein assay kit, by Thermo Fisher Scientific (2 mentions)
Hplc grade acetonitrile, by Merck Group (2 mentions)
Tween 80, by Merck Group (2 mentions)
Jaws 2 crl 11904, by American Type Culture Collection (2 mentions)
Triton x 100, by Merck Group (2 mentions)
Sybr green master mix, by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by ScienCell (1 mentions)
23 protocols using alpha minimum essential medium α mem
1
Fabrication of PCL-nHA Scaffolds for hBMSCs
2
Osteoclast Differentiation Assay
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DIO with a purity > 97% was purchased from Mansite (Chengdu, China) and prepared at a stock concentration of 100 mM in dimethylsulfoxide (DMSO). Alpha minimum essential medium (α-MEM) and fetal bovine serum (FBS) were obtained from ThermoFisher (Sydney, Australia). Recombinant mouse M-CSF was obtained from R&D Systems (Minneapolis, MN, USA). Recombinant GST-rRANKL was prepared as previously described (Xu et al., 2000) .
Antibodies against ERK, phospho-ERK, p38, phospho-p38, JNK, phospho-JNK, IκBα, c-Fos and β-actin were obtained from Abcam (Cambridge, MA, USA). Antibody specific for NFATc1 was obtained from Santa Cruz Biotechnology (Dallas, TX, USA). The cell viability kit (CCK-8) was obtained from Dojindo Molecular Technologies (Kumamoto, Japan).
Tartrate-resistant acid phosphatase (TRAcP) staining kit was purchased from Sigma-Aldrich (St. Louis, MO, USA).
Antibodies against ERK, phospho-ERK, p38, phospho-p38, JNK, phospho-JNK, IκBα, c-Fos and β-actin were obtained from Abcam (Cambridge, MA, USA). Antibody specific for NFATc1 was obtained from Santa Cruz Biotechnology (Dallas, TX, USA). The cell viability kit (CCK-8) was obtained from Dojindo Molecular Technologies (Kumamoto, Japan).
Tartrate-resistant acid phosphatase (TRAcP) staining kit was purchased from Sigma-Aldrich (St. Louis, MO, USA).
3
Gellan Gum Hydrogel Synthesis and Characterization
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Gellan gum (GG) hydrogels were prepared as previously described [21 (link)] with some modifications. Gelzan (Sigma-Aldrich, Saint Louis, MI, USA) was filtered through a sterile filtration system (Techno Plastic Products AG, Trasadingen, Switzerland), frozen in liquid nitrogen and lyophilized (LyoAlfa 10/15, Telstar, Terrassa, Spain) for three days to obtain sterile, dried and purified material. This was dissolved in deionized (DI) H2O to attain a final concentration of 0.5%, 0.75%, and 1.25% (w/v). After complete dissolution of the polymer at 90 °C under stirring, the temperature was lowered to 40 °C and Alpha Minimum Essential Medium (α-MEM) (Life Technologies, Paisley, UK) was added and then dispensed into a hydrophobic surface using a syringe/pipette for viscous solutions to produce spherical hydrogel particles. The size of the particles was given by the dispensed amount (1 to 50 µL). The injectability was assessed by coupling a 27 G needle in a syringe (1 mL) by loading the particles in a GG vehicle solution. Images of the particles were acquires using a Stereo Microscope + Lamp (Schott KL 200, Stemi 1000, Carl Zeiss, Oberkochen, Germany) combined with AxioVision software (version 4.8.2.0, Carl Zeiss, Oberkochen, Germany) which allows to measure hydrogel particles diameter.
4
Flavonoids Modulate Osteoblast Differentiation
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Total Drynariae Rhizoma flavonoids (DRTF) were purchased from Beijing Qihuang Pharmaceutical Manufacturing Co., Ltd. (National Medicine Permit No. Z20030007, number of production: 04080081, the content of DRTF ≥ 80%). Alpha-Minimum Essential Medium (α-MEM) was purchased from Life Technologies (USA). Fetal bovine serum (FBS) was obtained from ExCell Biology, Inc. (China). Ascorbic acid (AA), β–glycerophosphate (β-GP) and Alizarin Red S were purchased from Sigma-Aldrich (USA). Rat turnover markers ELISA kits were purchased from Biocalvin (Suzhou, China). RIPA lysis solution and a BCA protein assay kit were purchased from HEART biological technology Co., Ltd. (China). The RNAprep Pure Tissue Kit was purchased from TianGen Biotech Co., Ltd. (China). The PrimeScript™ RT reagent kit (Perfect Real Time) and SYBR Premix Ex Taq™ reagent were purchased from TAKARA Biotechnology (China). Anti-OPG antibody was purchased abcam (USA). Anti-ERα and anti-ERβ antibodies were purchased from R&D Systems (USA). Anti-GAPDH (Glyceraldehyde-3-phosphate dehydrogenase) antibody was obtained from Proteintech (China). Anti-RANKL and Horseradish peroxidase (HRP) conjugated antibodies were purchased from Ruiying bio (China).
5
Osteoclast Differentiation Signaling Pathways
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Alpha-minimum essential medium (α-MEM) and fetal bovine serum (FBS) were purchased from Life Technologies (Carlsbad, CA, USA). Recombinant human M-CSF was purchased from R&D Systems (Minneapolis, MN, USA), and GST-RANKL was purchased from Oriental Yeast Co., Ltd (Shiga, Japan). Anti-trimethyl-histone H3 lysine 4 and anti-trimethyl-histone H3 lysine 27 were from Active Motif (rabbit polyclonal antibody, 39159, Carlsbad, CA, USA) and Millipore (rabbit polyclonal antibody, 07–449, Billerica, MA, USA), respectively. Anti-NFATc1 antibody detects all DNA binding domain-containing NFATc1 splicing isoforms, but not the closely related NFATc2 [27 (link)]. Anti-FAK, anti-Pyk2 and anti-Src antibodies were from Cell Signaling Technology (rabbit polyclonal antibody; Beverly, MA, USA).
6
Osteoclastogenesis Induction Protocol
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Recombinant human M-CSF was purchased from R&D Systems (Minneapolis, MN, USA), and TGF-β and SB431542 were from Sigma-Aldrich (St Louis, MO, USA). GST-RANKL was purchased from Oriental Yeast Co., Ltd (Shiga, Japan). Alpha-minimum essential medium (α-MEM) and fetal bovine serum (FBS) were purchased from Life Technologies (Carlsbad, CA, USA). Smad2/3 antibody was purchased from BD Biosciences (Monoclonal antibody, Mouse, 610843, San Jose, CA, USA), anti-trimethyl-histone H3 lysine 4 was from Activemotif (Polyclonal antibody, Rabbit, 39159, Carlsbad, CA, USA), anti-trimethyl-histone H3 lysine 27 was from Millipore (Polyclonal antibody, Rabbit, 07–449, Billerica, MA, USA), anti-β-actin was from Sigma-Aldrich (Polyclonal antibody, Rabbit, A2066, St Louis, MO, USA).
7
Dendritic Cell Culture Protocol
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Acetonitrile (HPLC grade), glycerol, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), human lactoferrin, nile red dye (NR), phosphate buffered saline (PBS) tablets, poly(vinyl alcohol) (PVA, MW 9-10 KDa; 80%), trifluoroacetic acid (TFA, HPLC grade), Tween 80 ® and antibiotic/antimycotic (100X) solution were obtained from Sigma-Aldrich, UK. L-leucine was purchased from BioUltra, Sigma, UK. Tissue culture flasks (25 and 75 cm 2 ) with vented cap, 96-well flat bottom and U shaped plates, acetone, dimethyl sulfoxide (DMSO) and tetrahydrofuran were purchased from Fisher Scientific, UK. Alpha minimum essential medium (α-MEM) and granulocyte macrophage colony-stimulating factor (GM-CSF) was purchased from Life technologies, UK. Fetal calf serum (FCS) heat inactivated was purchased from Biosera UK. Micro BCA™ protein assay kit was purchased from Thermo Scientific, UK. Dendritic cell lines, JAWS II (CRL-11904™) were purchased from American type culture collection (ATCC). The biodegradable polymer, poly(glycerol adipate-co-ωpentadecalactone), PGA-co-PDL was synthesized as described previously (Thompson et al., 2006) .
8
Dendritic Cell Cytotoxicity Assay
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Acetonitrile (HPLC grade), glycerol, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), human lactoferrin, nile red dye (NR), phosphate buffered saline (PBS) tablets, poly(vinyl alcohol) (PVA, MW 9-10 KDa, 80%), trifluoroacetic acid (TFA, HPLC grade), Tween 80® and antibiotic/antimycotic (100X) solution were obtained from Sigma-Aldrich, UK. L-leucine (L-leu) was purchased from BioUltra, Sigma, UK. Tissue culture flasks (25 and 75 cm2) with vented cap, 96-well flat bottom and U shaped plates, acetone, dimethyl sulfoxide (DMSO) and tetrahydrofuran were purchased from Fisher Scientific, UK. Alpha minimum essential medium (α-MEM) and granulocyte macrophage colony-stimulating factor (GM-CSF) was purchased from Life technologies, UK. Fetal calf serum (FCS) heat inactivated was purchased from Biosera UK. Micro BCA™ protein assay kit was purchased from Thermo Scientific, UK. Dendritic cell lines, JAWS II (CRL-11904™) were purchased from American type culture collection (ATCC). The biodegradable polymer, poly(glycerol adipate-co-ω-pentadecalactone), PGA-co-PDL was synthesized as described previously (Thompson et al., 2006) .
9
Nepetin Modulates Osteoclast Differentiation
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Nepetin (also known as Eupafolin or 6‐Methoxyluteolin) was purchased from ChromaDex (Los Angeles, CA, USA) and dissolved in dimethyl sulphoxide (DMSO). Foetal bovine serum (FBS) and alpha‐minimum essential medium (α‐MEM) were obtained from Gibco (Thermo Fisher Scientific, Waltham, MA, USA). Receptor activator of nuclear factor‐κB ligand (RANKL) and mouse recombinant macrophage‐colony‐stimulating factor (M‐CSF) were procured from R&D Systems (Minneapolis, MN, USA). Commercial grade titanium (Ti) particles in powdered form were from Alfa Aesar (Haverhill, MA, USA). Specific primary antibodies were obtained from Cell Signaling Technology (Danvers, MA, USA) or Abcam (Cambridge, UK). The antibodies used included IKKα, IKKβ, phosphorylated (p)‐IKKα/β, p‐IκBα, p‐p65, p‐ERK, p65, p‐JNK, p‐p38, IκBα, ERK, JNK, p38, NFATc1, c‐Fos, TRAF6, Beclin 1, LC3‐I/II, K63‐linkage specific polyubiquitin, TRAF3 and β‐actin.
10
Isolation and Expansion of Synovial Fluid Mesenchymal Stromal Cells
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The macroscopic SFs were isolated from the diluted synovial fluid samples individually and washed three times with phosphate-buffered saline (PBS) in sterile Petri dishes. The fragments were then digested with 2.5 ml of 4 mg/ml type I collagenase (Gibco) for 2.5 hours at 37°C. The SFs were dispersed by pipetting until they were no longer visible. Cells were centrifuged at 450 g for 5 min, and after centrifugation, the supernatant was discarded. The cells were washed with PBS once and then centrifuged again. After the supernatant was discarded, the cells were plated in T25 flasks (Corning) with 3 ml complete culture medium [alpha minimum essential medium (α-MEM; Gibco) supplemented with 10% fetal bovine serum (Gibco)]. The cells were expanded in monolayer culture. Portions of the cells from each sample were mixed and expanded in monolayer culture for further experimentation. The remaining synovial fluid dilutions were used for collecting SFMSCs.
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