Puromycin dose response curve The puromycin dose response curves were performed by seeding 5103 Bge cells per well into 6-well plates and 2 days later on puromycin dihydrochloride (Existence Technology, USA) was added to the culture medium at final concentrations of 0.05, 0.5 and 5 g/ml. (mingsheng88.org)
Antibiotic concentration1
When creating stable cell lines, the optimal antibiotic concentration that is toxic to your cells will need to be determined by performing an antibiotic titration combined with cell density titration. (cellbiolabs.com)
Aminonucleoside1
Puromycin is an aminonucleoside antibiotic, derived from the Streptomyces alboniger bacterium, that causes premature chain termination during translation taking place in the ribosome. (wikipedia.org)
Plasmid1
The Bge cells were transfected with the plasmid pLNHX_cHS4_puroR (Supplementary Fig. invertebrate cell collection, Bge, from embryos of the snail were partially rescued when cultured in the presence of 5 g/ml of puromycin. (mingsheng88.org)
Plates containing puromycin are stable for 1 month when stored at 4 °C.[citation needed] Puromycin resistance in yeast can also be conferred through expression of the puromycin N-acetyl-transferase (pac) gene. (wikipedia.org)
Primary normal human tracheobronchial epithelial cells (NHBE) from 2 donors were tranduced with a lentiviral vector containing an Nrf2 luciferase reporter with puromycin resistance elements. (mattek.com)
Stably transduced cells were selected by puromycin resistance, expanded and cyropreserved to produce large pools of reporter-expressing cells which were then utilized to produce well differentiated organotypic airway epithelial models. (mattek.com)
Ampicillin and puromycin antibiotic resistance genes provide selection in bacterial or mammalian cells respectively. (sigmaaldrich.com)
Gene3
Deletion of the gene encoding the multidrug efflux pump Pdr5 sensitizes cells to puromycin. (wikipedia.org)
The particles contain a Cas9 gene driven by an EF1a promoter, along with a Puromycin selection marker. (magainin.com)
The expression of each introduced PPAR gene was investigated using the various concentrations of doxycycline in the culture media. (biomedcentral.com)
Blasticidin4
Human IgG antibody Laboratories manufactures the blasticidin concentration for tcell selection reagents distributed by Genprice. (magainin.com)
The Blasticidin Concentration For Tcell Selection reagent is RUO (Research Use Only) to test human serum or cell culture lab samples. (magainin.com)
Q: What final concentration of blasticidin and puromycin are necessary for making stable cell lines? (cellbiolabs.com)
The concentration range for puromycin and blasticidin is usually between 1-10 µg/mL. (cellbiolabs.com)
Stably1
Puromycin (Solarbio, Beijing, China) was made use of for screening stably transduced cells in the concentration array of 1 g/mL. (estrogen-receptor.com)
ELISA1
Also, serum of soluble E-selectin (sE-selectin) concentration in gastric cancer patients are detected by ELISA, but increasing only in gastric cancer patients with peritoneal metastasis [ 9 ]. (biomedcentral.com)
Soluble1
Puromycin is soluble in water (50 mg/ml) as colorless solution at 10 mg/ml. (wikipedia.org)
Cells11
Puromycin is stable for one year as solution when stored at -20 °C. The recommended dose as a selection agent in cell cultures is within a range of 1-10 μg/ml, although it can be toxic to eukaryotic cells at concentrations as low as 1 μg/ml. (wikipedia.org)
Puromycin acts quickly and can kill up to 99% of nonresistant cells within 2 days. (wikipedia.org)
As puromycin inhibits protein synthesis in eukaryotic cells, researchers were able to show that injections of this drug will result in both short-term as well as long-term memory loss in mice. (wikipedia.org)
Given that puromycin is supplied at 10 mg/ml in 20 mM HEPES buffer (pH 7.2 C 7.5), cells cultured in 10 M HEPES (but without puromycin) served as settings. (mingsheng88.org)
This product uses two chemical polymers that will form a complex with the virus and will settle onto the cells, increasing the local virus concentration at the cell surface. (cellbiolabs.com)
To do a kill curve, incubate parental cells with increasing concentrations of your antibiotic (see Table 1 for an starting point of what ranges to try) and observe for cell death. (imanislife.com)
For reporter cell lines with selection genes, I recommend maintaining the cells in antibiotic at a concentration that kills greater than 95% of parental cells. (imanislife.com)
To determine an appropriate concentration you can either guess and hope that your cells don't all die - if you guess right you just saved some time - or you can perform a kill curve on your transfected/transduced cells. (imanislife.com)
The HEK293T cells were infected with viral vector, screened by puromycin to select highly expressed cells. (bvsalud.org)
ATP7B R778L HepG2 cells were grown in DMEM supplemented with 10% fetal bovine serum, 2 mM L-Glutamine, and 2ug/ml puromycin (Invitrogen, Grand Island, NY, USA). (beingmedicos.com)
The stable HepG2 cells overexpressing R778L ATP7B were obtained by continuous screening with 2 µg/mL puromycin for 2 weeks and checked by PCR and western blot analyses. (beingmedicos.com)
Dose response experiments were performed to determine a range that spanned non-toxic to moderately toxic concentrations. (mattek.com)
Mammalian1
Lethal concentrations of puromycin are much higher for strains of Saccharomyces cerevisiae than mammalian cell lines. (wikipedia.org)
Serum1
DC-SIGNR (DC-SIGN-related protein, also known as L-SIGN, CD299) as a member of C-type lectin belonging to selectin is found high serum concentration in colon cancer patients [ 13 ]. (biomedcentral.com)
Selection5
But use of puromycin for E. coli selection requires precise pH adjustment and also depends on which strain is selected. (wikipedia.org)
For hassle-free selection and optimum results the use of specially modified puromycin is possible. (wikipedia.org)
Selection with puromycin is rapid (3-6 days) and the required doses are quite small (1-10 μg/mL), which make it fast and effective. (imanislife.com)
it works well, but requires longer selection times (10-14 days) and quite high concentrations (up to 2 mg/mL) for many cell lines. (imanislife.com)
Well, for selection I recommend using a higher concentration of antibiotic. (imanislife.com)
Protein3
Puromycin is an antibiotic protein synthesis inhibitor which causes premature chain termination during translation. (wikipedia.org)
In this reaction, a puromycin molecule is chemically attached to the end of an mRNA template, which is then translated into protein. (wikipedia.org)
On the contrary, treatment with the protein synthesis inhibitorscycloheximide, anisomycin, or puromycin as well as prolonged exposure to the RNA synthesis inhibitor actinomycin D mimicked the biochemical modulating effects of 6AN on cisplatin action. (aspetjournals.org)
High4
The rate of incorporation of histidine- 14 C into hot TCA-insoluble material was reduced by 30% at the low concentration and by 80-90% at the high concentration. (rupress.org)
If the drug was added after the "transition point," cell division and oral primordium formation were completed with only slight delay at the low concentration, and with considerable delay (in some cases complete arrest) at the high concentration. (rupress.org)
These observations indicate that basal levels of PNMT activity require that the medulla receive the very high concentrations of hydrocortisone or corticosterone that are available to it by virtue of its unique location within an envelope of adrenal cortex. (aspetjournals.org)
In methane fermentation of waste waters containing high concentrations of organic polymers, the hydrolytic activity relevant to each polymer is of paramount significance, in that polymer hydrolysis may become a rate-limiting step for the production of simpler bacterial substrates to be used in subsequent degradation steps. (fao.org)
Purification1
The establishment of lentiviral constructs with ATP7B R778L mutation mainly included packaging, concentration, and purification. (beingmedicos.com)
Expression1
We found that the expression of each PPAR subtype was tightly controlled by the concentration of doxycycline in these established cell lines. (biomedcentral.com)
Higher1
The concentrations of betulinic acid in the small intestine were higher than heart, lung, kidney, and liver. (bvsalud.org)
Range1
0.994 6) over a wide concentration range for all components. (bvsalud.org)
Form1
The puromycin can then form a covalent link to the growing peptide chain allowing the mRNA to be physically linked to its translational product. (wikipedia.org)
Culture1
Puromycin is used in cell biology as a selective agent in cell culture systems. (wikipedia.org)
Results1
The results showed that the optimal filter for the dopamine receptor agonist in this study was the cell seeding count of 7×104, and the effective concentration of the positive drug was 1-100 µmol·L-1. (bvsalud.org)
Activity1
The resotration by dexamethasone treatment of the PNMT activity in medullas of hypophysectomized animals is blocked by concurrent administration of actinomycin D or puromycin. (aspetjournals.org)