The protocol for using human samples was approved by the Ethics Committee of National Cancer Center/Cancer Hospital, Chinese Academy of Medical Sciences (NCC/CH, CAMS). Cell lines of HepG2 and Huh7 were purchased from ATCC, L02 from SIBS cell bank of Chinese Academy of Sciences. HBV envelop protein and HBV core protein were purchased from Jianan Biotechnology (Beijing, China). Palmitic acid (PA), fatty acid-free bovine serum albumin (FF-BSA), myriocin (Myr), C6-ceramide, thioglycolate and diethylnitrosamine (DEN) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Detailed information for preparation of these reagents is provided in Additional file 1 : Supplementary methods [15 (link), 20 (link)–24 (link)].
C6 ceramide
Manufactured by Merck Group
Sourced in United States
C6-ceramide is a synthetic lipid molecule that is commonly used in research applications. It serves as a tool for studying the role of ceramides in biological processes. C6-ceramide is a short-chain ceramide analog that can be utilized in various in vitro and cell-based experiments.
Automatically generated - may contain errors
Lab products found in correlation
Bacterial sphingomyelinase, by Merck Group (2 mentions)
Hepg2, by American Type Culture Collection (1 mentions)
Thioglycolate, by Merck Group (1 mentions)
Diethylnitrosamine den, by Merck Group (1 mentions)
Beas 2b cell line, by American Type Culture Collection (1 mentions)
Trizol, by Thermo Fisher Scientific (1 mentions)
Anti β actin, by Cell Signaling Technology (1 mentions)
Tb green mixture, by Takara Bio (1 mentions)
Anti jak2, by Cell Signaling Technology (1 mentions)
Chelerythrine chloride, by Merck Group (1 mentions)
Anti β actin, by Merck Group (1 mentions)
Bafilomycin a1, by Merck Group (1 mentions)
Anti lc3, by Cell Signaling Technology (1 mentions)
Prolong gold antifade mountant with dapi, by Thermo Fisher Scientific (1 mentions)
C17 sphingosine, by Avanti Polar Lipids (1 mentions)
Ceramides, by Avanti Polar Lipids (1 mentions)
Diaphot 300 microscope, by Nikon (1 mentions)
Fugene hd, by Promega (1 mentions)
Thapsigargin, by Merck Group (1 mentions)
C2 ceramide, by Merck Group (1 mentions)
8 protocols using c6 ceramide
1
Hepatocyte Lipotoxicity Protocol
2
Investigating JAK2/STAT3 Signaling in BEAS-2B Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
BEAS-2B cell lines were acquired from the American Type Culture Collection (ATCC, Rockville, MA, USA). C6-ceramide, ammonium pyrrolidinedithiocarbamate (PDTC), tyrphostin B42 (AG490), and 6-Nitrobenzo[b]thiophene-1, 1-dioxide (Stattic) were acquired from Sigma-Aldrich (St Louis, MO, USA). Dulbecco’s Modified Eagle’s Medium (DMEM) and Fetal bovine serum (FBS) were obtained from HyClone (Logan, UT, USA). Anti-JAK2, anti-phosphorylated JAK2 (p-JAK2), anti-STAT3, anti-phosphorylated STAT3 (p-STAT3), anti-CD68, and anti-β-actin antibodies were acquired from Cell Signaling Technology (Beverly, MA, USA). IRDye 680RD Goat anti-Rabbit and Goat anti-Mouse IgG (H + L) was acquired from LI-COR Biosciences (Lincoln, NE, USA). TRIzol was acquired from Invitrogen Corporation (Carlsbad, California, USA). TB Green Mixture was acquired from TaKaRa (Otsu, Shiga, Japan).
3
Intact Cell Shortening and Ca2+ Transients
4
Ceramide-Induced Autophagy Regulation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Tissue culture medium was obtained from Lonza. Palmitate, fatty-acid-free BSA, bafilomycin A1 and C6-ceramide were obtained from Sigma–Aldrich. FB1, sphingosine kinase inhibitor (SKI), fumonisin B1 (FB1), D,L-threo-1-phenyl-2-palmitoylamino-3-morpholinopropan-1-ol (PPMP), and sphingosine-1-phosphate were from Biomol. Apo-ONE® Homogenous Caspase-3/7 Assay kits were from Promega. All solvents were from Merck Eurolab or Fisher Scientific. Ceramides and C17-sphingosine were from Avanti Polar Lipids. Anti-LC3 (Cell Signaling, clone 2775, dilution 1/1000 for Western-blot; clone D11, dilution 1/500 for immunofluorescence), and anti-β-actin (Sigma-Aldrich, St. Louis, MO, USA, clone 8H10D10, dilution 1/5000) antibodies were from Sigma-Aldrich. Secondary alexa fluor antibodies and ProLong™ Gold Antifade Mountant with DAPI were from Invitrogen™, Waltham, MA, USA.
5
Erythrocyte Morphology and Function
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For all experiments except for the blood count, heparin blood was retrieved from the retrobulbar plexus of mice. For the blood count, EDTA blood was analyzed using an electronic hematology particle counter (type MDM 905 from Medical Diagnostics Marx; Butzbach, Germany) equipped with a photometric unit for haemoglobin determination. Plasma erythropoietin levels were determined using an immunoassay kit according to the manufacturer’s instructions (R&D Systems, Wiesbaden, Germany). Murine erythrocytes were isolated by being washed two times with Ringer solution containing (in mM): 125 NaCl, 5 KCl, 1 MgSO4, and 32 HEPES/NaOH (pH 7.4), 5 glucose, and 1 CaCl2. Where indicated, sucrose (550 mM), C6 ceramide (50 μM; Sigma) or bacterial sphingomyelinase (0.01 U/ml; Sigma) were added to the Ringer solution. May-Grünwald staining was used to examine changes in erythrocyte shape. Briefly, 20 μl of erythrocytes were smeared and fixed using methanol onto a glass slide, and stained with 5% Giemsa Azur-Eosin (Merck Millipore, Germany) in phosphate-buffered saline (in mM: 1.05 KH2PO4, 2.97 Na2HPO4, 155.2 NaCl) for 20 min. Subsequently, images were taken on a Nikon Diaphot 300 Microscope (Nikon Instruments, Germany).
6
Calcium Signaling Assay Reagents
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Thapsigargin, phorbol 12,13-dibutyrate, C2-ceramide and C6-ceramide and C2-dihydroceramide were purchased from Sigma-Aldrich. FuGENE HD was from Promega (Madison, USA), and TransIT/Jurkat was from Mirus Corporation (Madison, USA). mAb OKT3 and mAb 4G10 were from Thermo Fisher Scientific and A488-conjugated goat anti-mouse γ2b was from Invitrogen. The genetically encoded Ca2+ indicator RGECO-1 (Zhao et al., 2011 (link)) was purchased from Addgene (plasmid #32444; Cambridge, USA). A488-CTxB was from Invitrogen and TX-100 Surfact-Amps was from Thermo Fisher Scientific.
7
Thrombin-Induced Lipid Signaling Pathway
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
α-Thrombin (human plasma, ≥2,800 NIH units/mg protein) was purchased from Calbiochem (La Jolla, CA, USA) and Sigma-Aldrich (St. Louis, MO, USA). [9,10-3H(N)]palmitic acid, [5,6,7,8,9,11,12,14,15-3H(N)]arachidonic acid was purchased from Amersham Biosciences (Buckinghamshire, England, UK). MTT (Thiazolyl Blue Tetrazolium Bromide), C6-ceramide, bacterial sphingomyelinase, and 2’,7’-dichlorodihydrofluorescein diacetate (DCFH-DA) were purchased from Sigma-Aldrich. BAPTA-AM (1,2-bis-(o-Aminophenoxy)-ethane-N,N,N’,N’-tetraacetic acid, tetraacetoxymethyl ester) was purchased from BIOMOL Research Labs., Inc (Ann Arbor, MI, USA). Silica gel 60 thin-layer chromatography (TLC) plates were purchased from Merck (Darmstadt, Germany). For the protein assay, the Bradford reagent was purchased from Bio-Rad (Hercules, CA, USA). Plasticware for tissue culture was purchased from NUNC (Waltham, MA, USA). The cleaved caspase-3 ELISA kit was purchased from Cell Signaling Technology (Danvers, MA, USA). AA and Fumonisin B1 were purchased from Sigma-Aldrich. Trolox was obtained from Tocris (Minneapolis, MN, USA) and the Fluo-4 NW Calcium Assay Kit was purchased from Invitrogen (Carlsbad, CA, USA).
8
Regulation of CD44 by ERK1/2 in Glioma Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To address the mechanism of ERK1/2 regulation of CD44 in glioma cells, we used PDRG1 knockdown and negative control U118 cells for CD44 promoter activity analysis. Meanwhile, wildtype U118 and 293T cells were used for the analysis of the effect of ERK1/2 on CD44 promoter activity treated with MEK inhibitor U0126 (S1901; Beyotime Biotechnology) or ERK activator C6 ceramide (860506P; Sigma). Different groups of cell lines were seeded in 96‐well plates and were co‐transfected with pGL3‐CD44 promoter (19 122; Addgene) and pGL3‐TR (Promega). After transfection, cells were treated with U0126 (10 and 20 µM) or ceramide C6 (10 µg/mL) for 16 hours before being lysed. The control groups were set accordingly. CD44 luciferase activity was evaluated by the Dual Luciferase Assay System (Promega) after transfection for 48 hours.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!