A549 (lung carcinoma) cell line was purchased from National Centre for Cell Sciences, Pune. Human colon carcinoma HCT116 (p53+/+/p53−/−) and lung Carcinoma cell line H1299 (p53−/−) were purchased from ATCC, USA. The cell lines were cultured in DMEM or RPMI, supplemented with 10% heat-inactivated fetal bovine serum (Lonza, USA), L-glutamine (2 mM), sodium pyruvate (100 μg/ml), non-essential amino acids (100 μM), streptomycin (100 μg/ml), and penicillin (50 U/ml; Himedia, India). Primary antibodies used are p53 (DO-1; Santa Cruz, USA), α-GFP (Cell signaling, USA), α-Tubulin (Thermo Scientific, USA), GAPDH and Histone-3 (Biobharati Lifescience, India). Precision plus protein dual color standards (Bio-Rad) was used to detect the molecular weight of proteins in SDS-PAGE. Cyt-D, Lat-B, and Jas purchased from Calbiochem, USA was used at 5 μM, 1 μM, and 50 nM concentration respectively followed by Dox (1 μM, Sigma, USA) for 45 minutes. We made working stocks of all drugs (dimethyl sulfoxide for Cyt-D, Lat-B, and Jas; sterile H2O for Dox).
Non essential amino acids (neaa)
Manufactured by HiMedia
Sourced in United States
Non-essential amino acids are a class of amino acids that can be synthesized by the body and are not required to be obtained from the diet. They play a fundamental role in various biological processes and cellular functions.
Automatically generated - may contain errors
Lab products found in correlation
Hct116, by American Type Culture Collection (1 mentions)
H1299, by American Type Culture Collection (1 mentions)
α tubulin, by Thermo Fisher Scientific (1 mentions)
Heat inactivated fetal bovine serum, by Lonza (1 mentions)
Precision plus protein dual color standard, by Bio-Rad (1 mentions)
α gfp, by Cell Signaling Technology (1 mentions)
P53 do 1, by Santa Cruz Biotechnology (1 mentions)
Penicillin, by HiMedia (1 mentions)
L glutamine, by Lonza (1 mentions)
Streptomycin, by HiMedia (1 mentions)
Fetal bovine serum (fbs), by Merck Group (1 mentions)
Dulbecco modified eagle medium (dmem), by HiMedia (1 mentions)
2 protocols using non essential amino acids (neaa)
1
Cytoskeletal Dynamics in Cancer Cell Lines
2
MTT Cytotoxicity Assay Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The cells
were cultured using DMEM (Himedia) media containing 10% FBS (Sigma-Aldrich),
1% nonessential amino acids (Himedia), and 1% antibiotic and antimitotic
solution (Gibco). Cells were developed and incubated at 37 °C,
with 95% relative humidity, in a CO2 incubator in a T25
flask and monitored continuously using an inverted Olympus microscope.
When 80% confluency was achieved, it was trypsinized and used for
the MTT assay.47 (link) For the MTT assay, 24-well
plates were reserved and seeded with 1 × 104 cells
per well and incubated in a CO2 incubator. After 24 h of
incubation, the cells were treated with different drug concentrations
of 6.25, 12.5, 25, 50, 100, and 150 μM. They were incubated
for 24 h, and the cell viability was determined using MTT 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium
bromide. Then, the resultant suspension was kept on a microvibrator
for 10 min, and the absorbance was recorded at λ = 570 nm in
an ELISA plate reader. The experiment was also performed in triplicate.
Data were represented as the growth inhibition percentage, i.e., %
growth inhibition = 100 – [(AD × 100)/AB], where AD is
the measured absorbance in wells that contain samples and AB is the
measured absorbance for blank wells (cells with the medium and the
vehicle).
were cultured using DMEM (Himedia) media containing 10% FBS (Sigma-Aldrich),
1% nonessential amino acids (Himedia), and 1% antibiotic and antimitotic
solution (Gibco). Cells were developed and incubated at 37 °C,
with 95% relative humidity, in a CO2 incubator in a T25
flask and monitored continuously using an inverted Olympus microscope.
When 80% confluency was achieved, it was trypsinized and used for
the MTT assay.47 (link) For the MTT assay, 24-well
plates were reserved and seeded with 1 × 104 cells
per well and incubated in a CO2 incubator. After 24 h of
incubation, the cells were treated with different drug concentrations
of 6.25, 12.5, 25, 50, 100, and 150 μM. They were incubated
for 24 h, and the cell viability was determined using MTT 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium
bromide. Then, the resultant suspension was kept on a microvibrator
for 10 min, and the absorbance was recorded at λ = 570 nm in
an ELISA plate reader. The experiment was also performed in triplicate.
Data were represented as the growth inhibition percentage, i.e., %
growth inhibition = 100 – [(AD × 100)/AB], where AD is
the measured absorbance in wells that contain samples and AB is the
measured absorbance for blank wells (cells with the medium and the
vehicle).
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!