Gibco high-quality selection agents offer unique solutions for your research needs, such as dual selection and rapid stable cell line establishment.

View our full list of antibiotics and antimycotics. In addition, explore the use of antibiotics in cell culture.

Eukaryotic selection antibiotics

Selection antibioticMost common selection usageCommon working concentrationAvailable powder sizesAvailable liquid sizes
BlasticidinEukaryotic and bacteria1–20 µg/mL50 mg10 x 1 mL, 20 mL
Geneticin (G-418)Eukaryotic100–200 µg/mL bacteria
200–500 µg/mL mammalian cells
1 g, 5 g, 10 g, 25 g20 mL, 100 mL
Hygromycin BDual-selection experiments and eukaryotic200–500 µg/mL20 mL
Mycophenolic acidMammalian and bacteria25 µg/mL500 mg
PuromycinEukaryotic and bacteria0.2–5 µg/mL10 x 1 mL, 20 mL
ZeocinMammalian, insect, yeast, bacteria, and plants50–400 µg/mL8 x 1.25 mL, 50 mL

Bacterial selection antibiotics

Selection antibioticMost common selection usageCommon working concentrationAvailable powder sizesAvailable liquid sizes
Actinomycin DBacteria1 µg/mL5 mg, 10 mg
Ampicillin sodium saltBacteria10–25 µg/mL200 mg
BlasticidinEukaryotic and bacteria50–100 µg/mL50 mg10 x 1 mL, 20 mL
Carbenicillin, disodium saltAgrobacterium and E. coli100–500 µg/mL5 g
Kanamycin sulfateBacteria100 µg/mL5 g, 25 g100 mL
Mycophenolic acidMammalian and Bacteria25 µg/mL500 mg
Neomycin sulfateBacteria50 µg/mL100 g
Polymyxin B sulfateBacteria100 units/mL25 MU
PuromycinEukaryotic and bacteria0.2–5 µg/mL10 x 1 mL, 20 mL
Streptomycin sulfateBacteria50–100 µg/mL100 g
ZeocinMammalian, insect, yeast, bacteria, and plants75–400 µg/mL8 x 1.25 mL, 50 mL

Eukaryotic selection antibiotics

Selection antibioticMost common selection usageCommon working concentrationAvailable powder sizesAvailable liquid sizes
BlasticidinEukaryotic and bacteria1–20 µg/mL50 mg10 x 1 mL, 20 mL
Geneticin (G-418)Eukaryotic100–200 µg/mL bacteria
200–500 µg/mL mammalian cells
1 g, 5 g, 10 g, 25 g20 mL, 100 mL
Hygromycin BDual-selection experiments and eukaryotic200–500 µg/mL20 mL
Mycophenolic acidMammalian and bacteria25 µg/mL500 mg
PuromycinEukaryotic and bacteria0.2–5 µg/mL10 x 1 mL, 20 mL
ZeocinMammalian, insect, yeast, bacteria, and plants50–400 µg/mL8 x 1.25 mL, 50 mL

Bacterial selection antibiotics

Selection antibioticMost common selection usageCommon working concentrationAvailable powder sizesAvailable liquid sizes
Actinomycin DBacteria1 µg/mL5 mg, 10 mg
Ampicillin sodium saltBacteria10–25 µg/mL200 mg
BlasticidinEukaryotic and bacteria50–100 µg/mL50 mg10 x 1 mL, 20 mL
Carbenicillin, disodium saltAgrobacterium and E. coli100–500 µg/mL5 g
Kanamycin sulfateBacteria100 µg/mL5 g, 25 g100 mL
Mycophenolic acidMammalian and Bacteria25 µg/mL500 mg
Neomycin sulfateBacteria50 µg/mL100 g
Polymyxin B sulfateBacteria100 units/mL25 MU
PuromycinEukaryotic and bacteria0.2–5 µg/mL10 x 1 mL, 20 mL
Streptomycin sulfateBacteria50–100 µg/mL100 g
ZeocinMammalian, insect, yeast, bacteria, and plants75–400 µg/mL8 x 1.25 mL, 50 mL


G418, Geneticin

Commonly known as G418 or G-418, Gibco Geneticin reagent is an aminoglycoside related to gentamicin and is commonly used as a selective agent for eukaryotic cells. As an analog of neomycin sulfate, it interferes with the function of 80S ribosomes and protein synthesis in eukaryotic cells.

The dominant-acting resistance genes (neor) used on common mammalian expression vectors contain elements derived from either transposons Tn601 (903) or Tn5. When grown in medium containing Geneticin selection agent, stable colonies of mammalian cells expressing these resistance markers can be generated in 10 to 14 days.

Ordering information for G418, Geneticin

What's in your G418?

To be effective, selection agents need to be reliable, highly pure, and consistent in nature. Geneticin selective antibiotic, considered a reagent of choice for nearly two decades, is the only G-418 reagent that delivers these qualities to your mammalian cell cultures.

Compared to other manufacturers’ G-418 products, geneticin antibiotic exhibits higher purity, so you can use less and produce greater selection pressure with minimal toxicity from contaminants.

When evaluating G-418 products, consider more than just potency. Look at purity and working range. Taken together, these three characteristics provide a true evaluation of the effectiveness of your G-418.

Purity. The purity of geneticin selection agent is consistently greater than 90%, as determined by HPLC—far higher than any other supplier. This means:

  • 15–30% lower concentrations of geneticin agent can be used, as compared to other G-418 products, to achieve comparable selection results
  • Surviving clonal colonies may arise faster, and cells may appear healthier when using geneticin instead of lower purity G-418 products

Potency. Potency as a descriptive term is often used synonymously with purity, but potency as reported in QC data is not related to purity. The potency value is a measure of bacterial growth inhibition.

  • Gentamicins that commonly contaminate G-418 preparations from other suppliers will add to potency in bacterial assays yet have no effect on mammalian cell selectivity.
  • Other bacterial contaminants in impure G-418 may be toxic to mammalian cells, consequently lowering the ED50 value.

Therefore, comparing potency values between different suppliers is not a relevant specification for mammalian cell selection.

ED50 assay is a true measure of eukaryotic growth selectivity as determined on reference NIH3T3 cells. Higher purity generally translates to higher ED50 values.

  • Toxic impurities lower the ED50 value, which means a narrower working range of antibiotic selection
  • A consistent ED50 from a supplier assures performance reproducibility from lot-to-lot in mammalian cell culture applications

This means when purchasing a new lot of Geneticin, you do not need to re-optimize for ideal antibiotic concentration (assuming no other media alterations). The wide ranges of ED50 displayed in different lots from alternative G-418 suppliers may require you to re-optimize for each lot. Lot-to-lot consistency with Geneticin is sufficiently high that you will not notice any difference in selection performance between lots.

Purity. The purity of geneticin selection agent is consistently greater than 90%, as determined by HPLC—far higher than any other supplier. This means:

  • 15–30% lower concentrations of geneticin agent can be used, as compared to other G-418 products, to achieve comparable selection results
  • Surviving clonal colonies may arise faster, and cells may appear healthier when using geneticin instead of lower purity G-418 products

Potency. Potency as a descriptive term is often used synonymously with purity, but potency as reported in QC data is not related to purity. The potency value is a measure of bacterial growth inhibition.

  • Gentamicins that commonly contaminate G-418 preparations from other suppliers will add to potency in bacterial assays yet have no effect on mammalian cell selectivity.
  • Other bacterial contaminants in impure G-418 may be toxic to mammalian cells, consequently lowering the ED50 value.

Therefore, comparing potency values between different suppliers is not a relevant specification for mammalian cell selection.

ED50 assay is a true measure of eukaryotic growth selectivity as determined on reference NIH3T3 cells. Higher purity generally translates to higher ED50 values.

  • Toxic impurities lower the ED50 value, which means a narrower working range of antibiotic selection
  • A consistent ED50 from a supplier assures performance reproducibility from lot-to-lot in mammalian cell culture applications

This means when purchasing a new lot of Geneticin, you do not need to re-optimize for ideal antibiotic concentration (assuming no other media alterations). The wide ranges of ED50 displayed in different lots from alternative G-418 suppliers may require you to re-optimize for each lot. Lot-to-lot consistency with Geneticin is sufficiently high that you will not notice any difference in selection performance between lots.

SpecificationInvitrogenSupplier ASupplier BThe Geneticin Advantage

Purity (HPLC)

>90–93%

66–75% 

65–82%

Significantly higher purity

Potency claimed (µg/mg)

718–735

712–724

673–735

More stringent potency assay

Potency upon retest (µg/mg)

718–735

640–659

621–677

Consistently higher actual potency

ED50 Assay (µg/mL)

2,450–2,700

1,350–3,100

600–2,350

Reliable selection lot-to-lot

Table 1. Multiple lots of G-418 from two different suppliers were analyzed according to Invitrogen raw material QC protocols for geneticin. It was observed that results obtained with G-418 from Supplier A and Supplier B displayed wide variations on all measured parameters, including an inconsistency between the actual potency and the value reported on the supplied product literature. Discrepancies between re-tested potency values and that claimed by other suppliers may be due to differences in G-418 reference lots and assay procedures.
HPLC data of Geneticin compared to supplier A and supplier B

Figure 1. Comparison of HPLC data of geneticin with selected lots of G-418 from Supplier A and Supplier B. Samples were prepared and analyzed according to the procedure published in Clinical Chem. (1978), Vol. 24, No.11, p1940.


Zeocin

Gibco zeocin is highly effective in a variety of organisms, including mammalian and insect cell lines, as well as in yeast, bacteria, and plants. As a member of the bleomycin family, zeocin causes cell death by intercalating into and cleaving DNA.

Resistance to zeocin is conferred by the Sh ble gene product, which binds the antibiotic and prevents it from binding DNA. This selection agent is effective in multiple cell types, so eukaryotic expression vectors only need to carry one drug selection marker. This reduces the overall size of the vector and makes subcloning and transfection easier and more efficient.

Formula: C55H83N19O21S2Cu

Ordering information for zeocin

Cell typesConc. (μg/mL)*Invitrogen vectors carrying resistance marker
293 HEK200–400InsectSelect System, pIZ/V5-His vectors (stable expression in insect cells)
CHO~250pSecTag2, pSecTag2/Hygro (secreted mammalian expression)
COS-1~400ZeoCassette vectors (constructing Zeocin-resistant vectors)
HeLa<~150pcDNA4 vectors (constitutive mammalian expression)
Jurkat T cell~200pPICZ and pPICZ alpha vectors (inducible expression in Pichia pastoris)
NIH3T3~400T-REx System, pcDNA4/TO vectors (inducible mammalian expression)
Pichia pastoris~100 
S2 Drosophila~75 
Sf9 insect~250pGAPZ and pGAPZ alpha vectors (inducible expression in Pichia pastoris)

* The optimal concentration for selection of your cell line should be determined using kill curves.


Puromycin

Puromycin is an aminonucleoside antibiotic produced by the bacterium Streptomyces alboniger. During puromycin selection, this antibiotic inhibits cellular protein synthesis by disrupting peptide transfer on ribosomes causing premature chain termination during translation. It is a potent translational inhibitor in both prokaryotic and eukaryotic cells. Resistance to puromycin is conferred by the puromycin N-acetyltransferase gene (pac) from Streptomyces.

Puromycin selection at low concentrations

Puromycin has a fast mode of action, causing rapid cell death at low antibiotic concentrations. Adherent mammalian cells are sensitive to concentrations of 2 to 5 µg/mL, while cells in suspension are sensitive to concentrations as low as 0.5 to 2 µg/mL. Puromycin-resistant stable mammalian cell lines can be generated in less than one week.

Ordering information for puromycin

Cell lineConcentration [µg/mL]Reference
A549 (lung cancer)

1.5

Mol Cell Biol 27:324–339 (2007)
1.5J Biol Chem 283:33394–33405 (2008)
H1299 (non-small cell lung carcinoma)1Nucleic Acids Res 35:e17 (2007)
1.5J Biol Chem 283:33394–33405 (2008)
2.5PNAS 105:1937–1942 (2008)
HEK293 (human embryonic kidney)0.25Nucleic Acids Res 36:e83 (2008)
2Mol Cell Biol 28:4104–4115 (2008)
3Mol Cell Biol 27:4708–4719 (2007)
HeLa (human cervical cancer)1–2Genes Cells 12:397–406 (2007)
2Mol Cell Biol 28:4104–4115 (2008)
2PNAS 105:16490–16495 (2008)
Hep G2 (human hepatocellular carcinoma)0.3J Biol Chem 283:21462–21468 (2008)
2Mol Cell Biol 28:4104–4115 (2008)
2J Biol Chem 283:708–715 (2008)
HT1080 (human fibrosarcoma)0.2–0.4J Biol Chem 282:29314–29322 (2007)
1J Virol 82:3320–3328 (2008)
2Cancer Res 68: 1417–1426 (2008)
Human embryonic stem (ES) cells0.5Nucleic Acids Res 36:e148 (2008)
1Stem Cells 26:850–863 (2008)
5Stem Cells 26:2245–2256 (2008)

MCF-7 (human breast cancer)

0.45RNA 13:1375-1383 (2007)
1Cancer Res 68:1319–1328 (2008)
2Mol Cell Biol 28:4104–4115 (2008)
MDA-MB-231 (human breast cancer)0.5Mol Cell Biol 28:997–1006 (2008)
1.5Mol Cell Biol 27:324 – 339 (2007)
5Mol Cell Biol 28:687–704 (2008)


Blasticidin

Blasticidin is a nucleoside antibiotic produced by the bacterium Streptomyces griseochromogenes. It is a potent translational inhibitor in both prokaryotic and eukaryotic cells. Resistance to blasticidin is conferred by the product of the bsd gene from Aspergillus terreus.

Blasticidin selection at low concentrations

Blasticidin has a fast, potent mode of action, causing rapid cell death at low antibiotic concentrations. E. coli strains are generally sensitive to concentrations of 50 µg/mL, while mammalian cells are sensitive to concentrations as low as 2 to 10 µg/mL. Cell death occurs rapidly, and blasticidin-resistant stable mammalian cell lines can be generated in less than one week.

Ordering information for blasticidin

Cell LineConcentration [ug/mL]Reference
HT1080
(human fibrosarcoma)

1–5

10

20

J Biol Chem 283:11556–11564 (2008)

J Virol 82:7325–7335 (2008)

Mol Biol Cell 19:8–16 (2008)

HeLa
(human cervical cancer)

10

10

20

J Gen Virol 89:2611–2621 (2008)

Mol Biol Cell 19:8–16 (2008).

J Immunol 181:22–26(2008)

HEK293
(human embryonic kidney)

5

10

J Biol Chem 283:27534–27546 (2008)

J Virol 82:1665–1678 (2008)

Hep G2
(human hepatocellular carcinoma)

4

5

J Biol Chem 283:16320–16331 (2008)

Blood 113: 1786–1793 (2009)

HT1080
(human fibrosarcoma)

1–5

10

20

J Biol Chem 283:11556–11564 (2008)

J Virol 82:7325–7335 (2008)

Mol Biol Cell 19:8–16 (2008)

MCF-7
(human breast cancer)

2

5

Mol Cancer Res 6:555–567 (2008)

Mol Endocrinol 22: 361–379 (2008)

A549 (lung cancer)10Cancer Res 68:5040–5048 (2008)


Hygromycin B

Hygromycin B is an aminoglycosidic antibiotic that inhibits protein synthesis by disrupting translocation and promoting mistranslation at the 80S ribosome. Because it uses a different mode of action than geneticin, blasticidin S, or zeocin, it’s useful for dual-selection experiments when used in conjunction with another selection agent.

Hygromycin selection genes and concentrations

Resistance to hygromycin B is conferred by the E. coli hygromycin resistance gene (hyg or hph). The concentration for selection ranges from 100–1,000 µg/mL (typically 200 µg/mL) and should be optimized for each cell line.

Formula: C20H37N3O13
FW: 527.5 g/mole

Ordering information for hygromycin

Cell typesConc. (μg/mL)*Invitrogen vectors carrying resistance marker
CHO~250pIND/Hygro vector (Ecdysone-inducible mammalian expression)
HeLa~550pcDNA5 vectors (constitutive mammalian expression)
Jurkat T cell~1,000pSecTag2/Hygro vector (secreted mammalian expression)
S2 Drosophila200–300pREP vectors (episomal mammalian expression)
pCoHygro (selection vector for DES)

* The optimal concentration for selection of your cell line should be determined using kill curves.

For Research Use Only. Not for use in diagnostic procedures.

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