Human lung cancer A549 cells and human cervical carcinoma HeLa cells were grown in Roswell Park Memorial Institute (RMPI) 1640 medium without folic acid (Thermo Fisher Scientific), and human L-O2 hepatocytes were cultured in RMPI 1640 medium with folic acid (Thermo Fisher Scientific). All media were supplemented with 10% fetal bovine serum (FBS) (Thermo Fisher Scientific), and cells were maintained at 37°C and 5% CO2 in a humidified incubator. HeLa, A549, and L-O2 hepatocytes were provided by the Chinese Academy of Cell Resource Center (Shanghai Institute of Biological Sciences, Chinese Academy of Sciences, Shanghai, People’s Republic of China).
Folic acid
Manufactured by Thermo Fisher Scientific
Sourced in United States
Folic acid is a synthetic form of the B-vitamin folate. It is commonly used in laboratory settings as a nutritional supplement or as a component in various biological assays and cell culture media.
Automatically generated - may contain errors
Lab products found in correlation
L glutamine, by Thermo Fisher Scientific (5 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (5 mentions)
Inositol, by Merck Group (4 mentions)
Fetal bovine serum (fbs), by Merck Group (4 mentions)
Sodium bicarbonate, by Merck Group (4 mentions)
Horse serum, by Thermo Fisher Scientific (3 mentions)
Recombinant il 2, by Thermo Fisher Scientific (3 mentions)
Fetal bovine serum (fbs), by Corning (3 mentions)
Dulbecco modified eagle medium (dmem), by Corning (3 mentions)
Myo inositol, by Merck Group (3 mentions)
Rpmi 1640, by Merck Group (3 mentions)
2 mercaptoethanol, by Thermo Fisher Scientific (3 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
β mercaptoethanol, by Merck Group (2 mentions)
Alpha mem without nucleosides, by Thermo Fisher Scientific (2 mentions)
Penicillin streptomycin, by Merck Group (2 mentions)
Penicillin streptomycin, by Corning (2 mentions)
β mercaptoethanol, by Thermo Fisher Scientific (2 mentions)
Interleukin 2 (il 2), by Thermo Fisher Scientific (2 mentions)
Sodium bicarbonate, by Thermo Fisher Scientific (2 mentions)
23 protocols using folic acid
1
Cell Culture of Common Cancer Lines
2
Culturing Diverse Hematological Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
DC4 + human primary AML samples and normal peripheral blood mononuclear cells (PBMCs) were obtained from residual samples using a protocol approved by the Institutional Review Board of Stony Brook University. THP-1, U937, TALL104, and NK-92 cell lines were obtained from ATCC (Manassas, VA, USA). MOLM-13 was obtained from AddexBio (San Diego, CA, USA) T cells were cultured in filtered T cell media, defined as 50% AIM V, 40% RPMI 1640 and 10%FBS, with 1% Pen/Strep (all Gibco, Waltham, MA, USA) and supplemented with IL-2 (300 IU/mL; Peprotech, Rocky Hill, NJ, USA), unless otherwise specified. NK-92 cells were cultured in filtered NK cell media, defined as alpha-MEM without ribonucleosides and deoxyribonucleosides with 2mM L-glutamine, 1.5 g/L sodium bicarbonate (Gibco), 12.5% heat-inactivated horse serum (Gibco), 12.5% heat-inactivated FBS (Atlanta Biologicals, Atlanta GA, USA), 1% Pen/Strep (Gibco), 0.2% inositol (Sigma), 0.02% folic acid (Fisher), and 50 uM beta-mercaptoethanol (Fisher), supplemented with IL-2 (300 IU/mL), unless otherwise specified. THP-1, U937, and MOLM-13 cell lines were cultured in RPMI, 10% FBS, 1% Pen/Strep (Gibco). TALL104 cells were cultured in IMDM adding 300 IU/ml recombinant human IL-2, 2.5 mg/ml human albumin, 0.5 mg/ml D-mannitol, and 20% FBS.
3
Culturing Diverse Cell Lines and PBMCs
4
Characterization of NK92 Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
NK92, a human NK lymphoma cell line (CRL-2407) was obtained from ATCC. NK92 cells, expressing recombinant NKp46 or NKp46-IRES-GFP (designated as NK92.p46 or NK92.p46-IRES-GFP, respectively) were kindly provided by Kerry S Campbell (Fox Chase Cancer Center, Philadelphia, PA, USA). NK92 cell lines were grown in MEM Alpha medium (Gibco, Life Technologies), supplemented with heat-inactivated 10% horse serum, 10% FBS (Serum Source International), 0.2 mM myo-inositol (Sigma), 0.1 mM β-mercaptoethanol (Sigma) 0.02 mM folic acid (Fisher Scientific), 200 IU/mL of recombinant human IL-2 (eBioscience), and 50 IU/mL penicillin/streptomycin (Life Technologies) The following target cell lines were used HeLa, human cervical adenocarcinoma (ATCC CCL-2); HepG2, human hepatocellular carcinoma (ATCC HB-8065); 721.221, EBV-transformed human B-cell lymphoma. Cell lines were grown in a 5% CO2 humidified 37°C incubator and cultured in RPMI 1640 (Mediatech, Inc.) or DMEM (Gibco, Life Technologies) medium supplemented with 10% FBS and 1% penicillin/streptomycin. Antibodies that were used in this study were anti-human NKp46 PE or Biotin (Biolegend, clone 9E2), anti-human CD3 FITC (BD Pharmingen, clone UCHT1), and anti-human CD56 PE-Cy5 (BD Pharmingen, clone B159).
5
Pemetrexed Nanoparticle Synthesis Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Pemetrexed
disodium heptahydrate and poly-l -lysine hydrochloride (PLL;
15–30 kDa) were purchased
from Sigma Aldrich. Folic acid was purchased from Fisher Scientific.
All LbL methods were carried out in Milli-Q water from a Milli-Q source.
The KR2i TFF system was purchased from Spectrum Laboratories (Repligen,
Waltham, MA).
disodium heptahydrate and poly-
15–30 kDa) were purchased
from Sigma Aldrich. Folic acid was purchased from Fisher Scientific.
All LbL methods were carried out in Milli-Q water from a Milli-Q source.
The KR2i TFF system was purchased from Spectrum Laboratories (Repligen,
Waltham, MA).
6
Isolation and Culture of Primary Immune Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human peripheral blood mononuclear cells (PBMCs) were obtained from Immunospot and donors were uncharacterized with respect to sex or age. Primary NK and T cells were isolated from leukoreduction system chambers (LRS chambers) obtained from We Are Blood (Austin, TX); T cells were isolated from three male donors (ages 50, 58, and 71 years) and NK cells were isolated from two 49 year old female donors and a 57 year old male donor. Primary NK cells, primary T cells, HEK293T, Jurkat, and NK-92 cells were maintained at 37°C, 5% v/v CO2 in a humidified incubator. HEK293T cells were maintained in DMEM (Corning) supplemented with 10% FBS (Sigma) and 1% Penicillin-Streptomycin (Corning). Jurkat cells were cultured in RPMI 1640 (Sigma), 10% FBS (Sigma), 1% Penicillin-Streptomycin. NK-92 cells were cultured in NK media; Alpha MEM without nucleosides (GIBCO) with 2 mM L-glutamine (Fisher), 1.5 g/l sodium bicarbonate (Sigma), 0.2 mM inositol (Sigma), 0.1 mM 2-mercaptoethanol, 0.02 mM folic acid (Fisher), 100U/ml recombinant IL-2 (Peprotech), 12.5% horse serum (GIBCO) and 12.5% FBS (Corning).
7
Isolation and Culture of Primary Immune Cells
8
Quantitative Analysis of Vitamins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Thiamine hydrochloride (98.5–101.5%), Riboflavin (98%), Nicotinamide (98%), D-biotin (99%), Folic acid (99%) and cyanocobalamin (99%) were purchased from Fisher (Germany). Vitamins B5 (Pantothenic acid) and B6 (Pyridoxine hydrochloride) were purchased from Chem-Lab (Zedelgem, Belgium) and Fluorochem (United Kingdom) respectively. HPLC grade methanol, acetonitrile, chloroform (≥99.9%, containing amylenes as stabilizer) and salts for preparing the buffer solution were supplied by Sigma-Aldrich (Steinheim, Germany). Water used throughout the analyses was from a Millipore system.
9
Layer-by-Layer Preparation of Nanoparticles
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Raltitrexed, and poly-l -lysine hydrochloride (PLL, 15–30 kDa) were purchased from Sigma Aldrich. Folic acid was purchased from Fisher Scientific. All LbL methods were carried out in Milli-Q water from a Milli-Q source. KR2i TFF system was purchased from Spectrum Laboratories (Repligen, Waltham, MA, USA).
10
Generating NKp46 Isoform Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
NKp46 full length, i.e., domain 1+ (isoform a; matching NM_004829.5) and NKp46-D2, i.e., domain 1− (isoform d; matching NM_001242356.1) cDNA were N-terminal FLAG-tagged and cloned into pBMN-IRES-GFP retroviral vector. NK-92 cells (ATCC CRL-2407), derived from human NK cell leukemia were then transduced with the constructs to produce NK-92-NKp46-Full (NK92-46Full) and NK-92-NKp46-D2 (NK92-46D2) cell lines, respectively. Retroviral transduction was performed as previously described (21 (link)–24 (link)). NK-92 cell lines were then grown in Alpha-MEM medium (Gibco, Life Technologies) supplemented with 10% horse serum, 10% FBS, 0.2 mM myo-inositol (Sigma), 0.1 mM β-mercaptoethanol (Sigma), 0.02 mM folic acid (Fisher Scientific), 100 IU/mL of recombinant human IL-2 (PeproTech), and 1% penicillin/streptomycin (Life Technologies). HEK293T, SV40 large T Ag-transfected HEK293 cells (CRL-11268) were used as target cell lines. HEK293T cells were cultured in DMEM (Gibco, Life Technologies) medium supplemented with 10% FBS and 1% penicillin/streptomycin. Cell lines were grown in a 5% CO2 humidified 37°C incubator.
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!