Cell Culture Antibiotic Selection Guide

• Why use antibiotics in cell culture?
• Antibiotic kill curve
• Antibiotics for Mammalian Cell Culture
• Antibiotics for Plant Cell Culture
• Antibiotic Selection Agents for Cell Culture

Antibiotics are commonly used in cell culture to prevent contamination, maintain aseptic conditions, or select for cells containing genetic modifications. The use of antibiotics can minimize loss of cell lines and cell cultures, saving time and conserving precious resources. Proper antibiotic selection depends on both cell type and which contaminants you are trying to prevent. Selective antibiotics for generating stable cell lines or other recombinant cultures should be chosen based on the antibiotic resistance gene or selectable marker. The tables below can be used to guide antibiotic selection for your cell culture needs.

Antibiotics for Mammalian Cell Culture

† = Effective against most species
‡ = Effective against certain species

Product Name	Product No.	Gram (+)bacteria	Gram (-)bacteria	Yeasts	Molds	Mycoplasma	Suggested Working Conc.
Amphotericin B	A2411			†	†		2.5 mg/L
Amphotericin B-Solubilized (Approx. 45%)	A9528			†	†		5.6 mg/L (of solid)
Amphotericin B(250 μg/ml solution)	A2942			†	†		10 ml/L
Ampicillin	A0166	†	†				100 mg/L
Antibiotic Antimycotic Solution (100x)(10,000 units penicillin, 10 mg streptomycin, and 25 µg amphotericin B per ml)	A5955	†	†	†	†		10 ml/L
Cephalothin	C4520	†	†				100 mg/L
Dihydrostreptomycin	D5155	†	†				100 mg/L
Erythromycin	E5389	†	†				100 mg/L
Gentamicin Sulfate	G1264	†	†			†	50 mg/L
Gentamicin Sulfate (10 mg/ml solution)	G1272	†	†			†	5 ml/L
Gentamicin Sulfate (50 mg/ml solution)	G1397	†	†			†	1 ml/L
L-Glutamine-Penicillin-Streptomycin Solution (200 mM L- glutamine, 10,000 units penicillin, and 10 mg streptomycinper ml)	G1146  G6784	†	†				10 ml/L
Kanamycin Sulfate (10 mg/ml solution)	K0129	†	†			†	10 ml/L
Kanamycin Sulfate (50 mg/ml solution)	K0254	†	†			†	2 ml/L
Lincomycin HCI	L2774	†					100 mg/L
Neomycin Sulfate	N6386	†	†				50 mg/L
Neomycin Sulfate (10 mg/ml solution)	N1142	†	†				5 ml/L
Nystatin (5,000 units nystatin per mg)	N6261			†	†		2.5 X 105U/L (50 mg/L)
Nystatin	N4014			†	†		50 mg/L
Nystatin Suspension	N1638			†	†		24 ml/L
Paromomycin Sulfate	P5057	†	†				100mg mg/L
Penicillin-G (potassium salt)	P7794	†					100,00 U/L
Penicillin-G (sodium salt)	P3032	†					100,000 U/L
Penicillin-Streptomycin solution (5,000 units penicillin-G and 5 mg streptomycin per ml)	P4458	†	†				20 ml/L
Penicillin-Streptomycin Solution (10,000 units penicillin-G and 10 mg streptomycin per ml)	P0781  P7539  P4333	†	†				10 mg/L
Penicillin-Streptomycin-Neomycin solution (5,000 units penicillin-G,5 mg streptomycin and 10 mg neomycin per ml)	P4083	†	†				10 ml/L
Phenoxymethylpenicillinic Acid (potassium salt) [Penicillin V]	P4807	†					100,000 U/L
Polymyxin B Sulfate	P4932		†				50 mg/L
Spectinomycin dihydrochloride pentahydrate	S4014	†	†				7.5-20 mg/L
Streptomycin sulfate	S9137	†	†				10 mg/L 100 μg/ml
Tetracycline Hydrochloride	T7660	†	†				10 mg/L
Tylosin Tartrate(8 mg/ml soln.)	T3397	†				†	1 ml/L

Antibiotics for Plant Cell Culture

Product Name	Product No.	Gram (+)bacteria	Gram (-)bacteria	Mycobacteria	Yeasts	Molds	Mycoplasma	Suggested Working Conc.
Carbenicillin	C3416	‡	†					10-500 μg/ml
Cefotaxime	C7039	‡	†					up to 500 μg/ml (CC, PR, EC)
Chloramphenicol	C1919	†	†	‡			‡	10-35 μg/ml (TS)
G 418	G1279							up to 250 μg/ml (CC)
Gentamicin Sulfate	G1264	‡	†				†	up to 500 μg/ml (CC, PR, EC)
Hygromycin B	H9773							up to 500 μg/ml (CC, PR, EC)
Kanamycin	K1377	†	†				†	16 (SSC); up to 40 μg/ml (CC, PR, EC)
Paromycin	P8692	†	‡
Penicillin G, Potassium	P7794	†	‡					up to 100 μg/ml
Penicillin G, Sodium	P8431	†	‡					up to 100 μg/ml
Pentachloronitrobenzene	P8556					†
Rifampicin	R7382	†	†	†				50 (TS); up to 25 μg/ml (PC, TEC)
Thiabendazole	T5535					†
Vancomycin	V1130	†						up to 10 μg/ml (PC); up to 40 μg/m (CC, EC, STM)

† = Effective against most species
‡ = Effective against certain species

¹Application Key: CC = Callus Culture; PR = plant regeneration; EC = cotyledon, hypocotyl, or leaf disc culture; TS = transformant
selection; SE = somatic embryogenesis; SSC = stem selection culture; PC = protoplast culture; TEC = tuber explant culture;
STM = shoot tip micropropagation

Antibiotic Selection Agents for Cell Culture

Product Name	Product No.	Mode of Action	Suggested Working Conc.
Actinomycin D	A9415	Complexes with DNA and interferes with RNA synthesis	1 µg/ml
Bleomycin Sulfate	B8416	Complexes with DNA, causing strand scissions	10-100 µg/ml
Chloramphenicol	C3175	Inhibits elongation at peptidyl transferase	5 µg/ml
G 418	A1720	Blocks polypeptide synthesis and inhibits chain elongation	100-800 µg/ml
G 418 (50 mg/ml solution)	G8168	Blocks polypeptide synthesis and inhibits chain elongation	1 µg/ml
Hygromycin B	H3274	Blocks polypeptide synthesis and inhibits chain elongation
Mitomycin C	M4287	Inhibits nucleic acid synthesis	10-50 µg/ml
Mycophenolic Acid	M3536	Blocks inosine monophosphate dehydrogenase in guanosine monophosphate pathway	25 µg/ml
Puromycin HCl	P8833	Inhibits protein synthesis	10-100 µg/ml