How do you dilute Blasticidin?
How do you dilute Blasticidin?
Preparing and Storing Stock Solutions
- Prepare Blasticidin stock solutions of 5-10 mg/ml by dissolving Blasticidin in sterile water and filter-sterilize the solution.
- Aliquot in small volumes suitable for one time use and store at 4ºC (short term) or at -20ºC (long-term).
How long is Blasticidin stable?
Blasticidin solution is stable for 9 months at −20°C to −5°C. Medium containing Blasticidin can be stored at 4°C for up to 2 weeks. On thawing, use what you need and discard the unused portion. For selection of Blasticidin-resistant E.
How long is Blasticidin selection?
Cell death induced by blasticidin occurs rapidly, allowing for selection of transfected cell lines carrying a blasticidin resistance gene within one week.
How do you make 5% saponin?
Saponin Stock Solution (10x, 5% w/v): To create a 5% w/v solution (10x), dissolve 5 grams of Saponin (Sigma 47036-50g-F) into 80ml water. Mix thoroughly until dissolved by rocking overnight at 4 deg C. q.v. 100ml, filter sterilize through 0.2 um filter, and store at 4 deg C.
How do you make a 10% saponin?
10% Saponin stock: Mix 5g Saponin (Sigma #S-7900) in 50 ml CMF-PBS. Place at 37˚C until the saponin has dissolved completely. Use a 50 ml syringe with 0.2μm disposable filter cartridge to filter the solution.
How do you make Blasticidin stock solution?
Stock solutions guidelines Prepare Blasticidin stock solutions of 5–10 mg/mL by dissolving Blasticidin in sterile water and filter-sterilize the solution. Blasticidin is soluble in water and acetic acid. Aliquot in small volumes appropriate for one time use and store at 4°C (short term) or at −20°C (long-term).
How do you make a saponin solution?
Saponin permeating solution is made by mixing and dissolving 0.5g of Saponin with 100ml of 10X PBS, followed by submicron filtration and/or autoclaving at 15 PSI for 60 minutes.
How do you dilute saponin?
Antibody Dilution Buffer (1X PBS / 1% BSA / 0.1% Saponin): To prepare 40 ml, add 4 ml 10X PBS to 36 ml dH2O, mix. Add 0.4 g BSA and mix well. While stirring, add Saponin to final concentration of 0.1%.
How does puromycin inhibit protein synthesis?
Puromycin is a naturally occurring aminonucleoside antibiotic that inhibits protein synthesis by ribosome-catalyzed incorporation into the C-terminus of elongating nascent chains, blocking further extension and resulting in premature termination of translation.
How do you make a 5% saponin solution?
To create a 5% w/v solution (10x), dissolve 5 grams of Saponin (Sigma 47036-50g-F) into 80ml water. Mix thoroughly until dissolved by rocking overnight at 4 deg C. q.v. 100ml, filter sterilize through 0.2 um filter, and store at 4 deg C.
How do you prepare blasticidin stock solutions?
Prepare Blasticidin stock solutions of 5–10 mg/mL by dissolving Blasticidin in sterile water and filter sterilize the solution. Blasticidin is soluble in water and acetic acid. Aqueous stock solutions are stable for 1–2 weeks at 4°C and 6–8 weeks at −20°C.
What is blasticidin S used to treat?
Blasticidin S is a nucleoside antibiotic isolated from Streptomyces griseochromogenes. It is a potent inhibitor of protein synthesis in both prokaryotic and eukaryotic cells. Blasticidin S acts quickly by inhibiting peptide bond formation by the ribosome. Blasticidin S is used for the prevention of contamination of cell cultures.
How much blasticidin does it take to kill cells?
Typically, mammalian cells are sensitive to blasticidin concentrations of 1-10 µg/ml, and bacteria to 25-100 µg/ml. Cell death induced by blasticidin occurs rapidly, allowing for selection of transfected cell lines carrying a blasticidin resistance gene within one week.
What is blasticidin-S deaminase resistance?
Resistance to blasticidin is conferred by the blasticidin resistance gene from Bacillus cereus (bsr), which codes for blasticidin-S deaminase. Resistance to blasticidin is also conferred by the blastcidin S acetyltransferase gene (bls) from Streptoverticillum sp, and the blasticidin S deaminase gene (BSD) from Apergillus terreus.
https://www.youtube.com/watch?v=k9BEIFAAooY
