HT-29 (human colorectal adenocarcinoma) cells and MCF7 cells (human breast adenocarcinoma) were obtained from the ATCC (Manassas, VA). The MCF7 cells were cultured in Dulbecco’s modified Eagle medium (DMEM, GIBCO, Waltham, MA) supplemented with 10% fetal bovine serum (FBS) and 1 × antibiotics (GIBCO). The HT-29 cells were maintained in RPMI-1640 (GIBCO) medium supplemented with 10% FBS and 1 × antibiotics (GIBCO). Fatty acids were obtained from Nu-Chek Prep, Inc. (Elysian, MN); the BSA-bound Fatty acids were prepared as described in a previous study (Moon et al. 2001 (link)) and added to the culture medium. Delipidated FBS (DLFBS) was prepared as previously described (Hannah et al. 2001 (link)). The cultures were incubated in a humidified atmosphere of 5% CO2 at 37°C. Cerulenin (Sigma, St. Louis, MO) was dissolved in ethanol and added to the medium at 12.5 µM.
Fatty acids
Manufactured by Nu-Chek Prep
Sourced in United States
Fatty acids are a type of lipid compound that are essential for various biological functions. They are composed of a long hydrocarbon chain and a carboxyl group. Fatty acids serve as a source of energy and are important structural components of cell membranes.
Automatically generated - may contain errors
Lab products found in correlation
Deuterium oxide, by Cambridge Isotopes (1 mentions)
Tmr dextran, by Thermo Fisher Scientific (1 mentions)
Bafilomycin a, by Enzo Life Sciences (1 mentions)
Ultrapure e coli lps, by InvivoGen (1 mentions)
Lysotracker red, by Thermo Fisher Scientific (1 mentions)
Hplc grade, by Merck Group (1 mentions)
T7 endonuclease, by New England Biolabs (1 mentions)
Hp taq dna polymerase, by Bio Basic (1 mentions)
Q5 dna polymerase, by New England Biolabs (1 mentions)
Restriction enzyme, by New England Biolabs (1 mentions)
Fatty acid free bsa, by MP Biomedicals (1 mentions)
1 14c fatty acids, by PerkinElmer (1 mentions)
Cerulenin, by Merck Group (1 mentions)
Mcf 7 cell, by American Type Culture Collection (1 mentions)
Mcf 7, by Thermo Fisher Scientific (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Boron trifluoride, by Merck Group (1 mentions)
Dimyristoyl phosphatidylserine dmps, by Avanti Polar Lipids (1 mentions)
12 protocols using fatty acids
1
Culturing Colorectal and Breast Cancer Cells
2
Fatty Acid Analytical Standards
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Fatty acids used in this study were purchased from Nu Chek Prep, Inc. (MN, USA). Deuterium oxide (99.9%) was purchased from Cambridge Isotope Laboratories (Tewksbury, MA, USA). All other solvents and chemicals were reagent grade or better and were used as purchased without further purification.
3
Lysosome Dynamics and Macrophage Function
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
CAO74-ME and bafilomycin A were from Enzo Life Sciences. Lysotracker red and TMR-dextran (10,000 MW) were from Invitrogen. The cathepsin B activity assay was from Immunocytochemistry Technologies. Ultrapure E. coli LPS was from Invivogen. Thioglycollate was from Difco. Fatty acids were from Nu-Chek Prep. The cathepsin D and LAMP1 antibodies were from Abcam. The actin antibody was from Sigma-Aldrich. The CD107a (LAMP-1) PE conjugated antibody was from eBiosciences (cat#12-1071). The ATG5 antibody was from Novus Biologics. Ultrapure-bovine serum albumin (BSA) was from Lampire and was tested for TLR ligand contamination prior to use.
4
Fatty acid procurement and cell culture
5
CRISPR/Cas9 System Reagents
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Agrobacterium and E. coli media were purchased from Bio Basic Inc. (York, Ontario, Canada). Fatty acids and their standards were obtained from Nu-Chek-Prep, Inc. (Elysian, MN, USA). Q5 DNA polymerase, restriction enzymes, T7 Endonuclease and deoxynucleotide triphosphate (dNTP) were purchased from New England Biolabs (Ipswich, MA, USA). HP Taq DNA polymerase was obtained from Bio Basic Inc. (York, Ontario, Canada). Primers were synthesized from Sigma-Aldrich (St. Louis, MO, USA). GC grade solvents were from Fisher Scientific (Ottawa, ON, Canada). All other chemicals were purchased from Sigma-Aldrich Ltd (Oakville, ON, Canada). The intermediate vector and expression vector for CRISPR/Cas9 system were purchased from Addgene (Watertown, MA, USA).
6
Preparation of Fatty Acid-BSA Conjugates
7
Fatty Acid Synthesis and Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Fatty acids and Coenzyme A were obtained from Nu-Chek Prep Inc (Elysian, MN) and from Larodan (Malmö, Sweden), [1-14C]Fatty acids from PerkinElmer or Lipidox (Stockholm, Sweden). Palmitoyl(16:0)-lysophosphatidylcholine (16:0-lysoPtdCho), essentially fatty acid free bovine serum albumin (BSA), and other fine chemicals were purchased from Sigma-Aldrich (St. Louis, USA). [14C]Acyl-CoA and acyl-CoAs were synthesized according to the method described by [19 (link)]. Argentation TLC plates were prepared by immersing TLC plates (silica 60, Merck) in AgNO3 (12%, w/v) dissolved acetone/water (4:1 by vol.) and drying the plates at room temperature.
8
Fatty Acid Procurement and Characterization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Fatty acids used in this study were purchased from Nu Chek Prep, Inc. (Elysian, MN, USA). All other solvents and chemicals were reagent-grade or better and were used as purchased without further purification.
9
Sourcing Chemicals for Research
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Fatty acids were purchased from Nu-Chek prep, Inc (Elysian, MN, USA). Other chemicals were purchased from Sigma-Aldrich (St. Louis, MO, USA), Invitrogen (Carlsbad, CA, USA), Calbiocem (La Jolla, CA, USA), Gibco (Grand Island, NY, USA) or Applied Biosystems (Foster City, CA, USA).
10
Fatty Acid Supplementation in C. elegans
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Fatty acids were obtained from Nu-Chek-Prep, Inc. and were prepared as previously described [42 ]. Assays were performed by growing synchronized L1 worms on the indicated fatty acid or control media containing tergitol (0.1%). Unless otherwise indicated, oleate was used at a concentration of 400 or 500 μM, except for Fig 1E , which used 1 mM. Palmitoleic acid and linoleic acid were used at a concentration of 500 μM.
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!