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Gibco™ Puromycin Dihydrochloride
Description
Puromycin works by inhibiting peptidyl transfer on both prokaryotic and eukaryotic ribosomes; resistance is conferred by the expression of the pac gene.
- Used in cell biology as selection antibiotic in mammalian cell culture systems
- Recommended working concentration ranges from 0.2–5.0μg/mL
- Can be toxic to eukaryotic cells at concentrations as low as 1μg/mL
- Supplied at 10mg/mL in 20 mM HEPES buffer (pH 7.2–7.5)
Cell Culture, Cloning, Drosophila S2 Cell Culture, Insect Cell Culture, Mammalian Cell Culture, Selection, Sf9 & Sf21 Cell Culture, Transfection, Transformation
Order Info
Shipping Condition: Dry Ice
Specifications
Specifications
| Content And Storage | Storage conditions: -5 to -20°C Shipping conditions: Frozen Shelf life: 12 months from date of manufacture |
| Concentration | 10 mg/mL |
| Culture Type | Mammalian Cell Culture, Insect Cell Culture |
| Form | Liquid |
| Product Type | Antibiotic |
| Sterility | Sterile-filtered |
| With Additives | HEPES |
| For Use With (Application) | Eukaryotic Selection⁄Stable Cell Line Generation |
| Product Line | Gibco |
| Quantity | 10 x 1 mL |
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Frequently Asked Questions (FAQs)
All of our antibiotics (Geneticin, Zeocin, Hygromycin B, Blasticidin, and Puromycin) can be used together for making multiple stable cell lines. However, kill curves will need to be performed for each combination of antibiotics since sensitivity to a given antibiotic tends to increase when combined with other antibiotics.
Puromycin Dihydrochloride is light sensitive on par with the light sensitivity of most basal media like DMEM and RPMI 1640. We would recommend limiting exposure of this product to light as much as possible (i.e,. don't leave on the bench or under hood lights longer than necessary). That said, using a light microscope to observe cells under normal conditions and timeframes will not break down the antibiotic.
When an irreplaceable culture becomes contaminated, researchers may attempt to eliminate or control the contamination.
1. Determine if the contamination is bacteria, fungus, mycoplasma, or yeast. Read more here to view characteristics of each contaminant.
2. Isolate the contaminated culture from other cell lines.
3. Clean incubators and laminar flow hoods with a laboratory disinfectant, and check HEPA filters.
4. Antibiotics and antimycotics at high concentrations can be toxic to some cell lines. Therefore, perform a dose-response test to determine the level at which an antibiotic or antimycotic becomes toxic. This is particularly important when using an antimycotic such as Gibco Fungizone reagent or an antibiotic such as tylosin.
The following is a suggested procedure for determining toxicity levels and decontaminating cultures:
1. Dissociate, count, and dilute the cells in antibiotic-free media. Dilute the cells to the concentration used for regular cell passage.
2. Dispense the cell suspension into a multiwell culture plate or several small flasks. Add the antibiotic of choice to each well in a range of concentrations. For example, we suggest the following concentrations for Gibco Fungizone reagent: 0.25, 0.50, 1.0, 2.0, 4.0, and 8.0 µg/mL.
3. Observe the cells daily for signs of toxicity such as sloughing, appearance of vacuoles, decrease in confluency, and rounding.
4. When the toxic antibiotic level has been determined, culture the cells for two to three passages using the antibiotic at a concentration one- to two-fold lower than the toxic concentration.
5. Culture the cells for one passage in antibiotic-free media.
6. Repeat step 4.
7. Culture the cells in antibiotic-free medium for four to six passages to determine if the contamination has been eliminated.
Please view the following page to browse the cell culture antibiotics we offer (https://www.thermofisher.com/us/en/home/life-science/cell-culture/mammalian-cell-culture/antibiotics.html).
For Research Use Only. Not for use in diagnostic procedures.
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