Ampicillin

Manufactured by AppliChem

Sourced in Germany

Ampicillin is a broad-spectrum antibiotic that belongs to the penicillin class. It is commonly used in laboratory settings for various applications, such as bacterial culture and selection. Ampicillin inhibits the synthesis of the bacterial cell wall, which is essential for the survival and growth of certain microorganisms.

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Lab products found in correlation

Kanamycin, by AppliChem (15 mentions) Chloramphenicol, by AppliChem (11 mentions) Streptomycin, by AppliChem (9 mentions) Kanamycin, by Carl Roth (6 mentions) Chloramphenicol, by Merck Group (6 mentions) Gentamycin, by AppliChem (5 mentions) Tetracycline, by AppliChem (4 mentions) M9 minimal medium, by BD (4 mentions) L cysteine, by Merck Group (3 mentions) Ciprofloxacin, by Merck Group (3 mentions) Erythromycin, by Merck Group (2 mentions) M9 minimal medium, by AppliChem (2 mentions) Gentamicin, by AppliChem (2 mentions) Phosphate buffered saline (pbs), by BioConcept (2 mentions) Horseradish peroxidase hrp conjugated rabbit anti sheep, by Jackson ImmunoResearch (2 mentions) Penicillin, by Thermo Fisher Scientific (2 mentions) Streptomycin, by Thermo Fisher Scientific (2 mentions) Nystatin, by AppliChem (2 mentions) Hemin chloride, by Merck Group (2 mentions) Non essential amino acids (neaa), by Thermo Fisher Scientific (2 mentions)

Spelling variants of this product

Ampicillin (Ap, (2 citations)

53 protocols using ampicillin

Standardized Pre-culture Procedure for Lactococcus lactis

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L. lactis MG1363^{37 (link)} and L. lactis MG1363_GFP were cultured in M17 broth supplemented with 2.0 wt% glucose, unless otherwise indicated. M17 supplemented with 1.0 wt% glucose and 1.1 wt% agarose was used for plate counting. Cultures and plates were incubated at 30 °C. As suggested by Harms et al. we standardized the pre-culture procedure (Fig. 1) and used exponential phase pre-cultures in balanced growth (13 generations in excess glucose)^{38 (link)}. 10 or 100 µg/mL ampicillin (Applichem, Darmstadt, Germany) was directly added to exponential pre-cultures; stationary pre-cultures were diluted 1/100 in fresh medium with ampicillin. Heat-killed cells were prepared by incubating stationary pre-cultures in phosphate buffered saline (PBS) of pH 7.5 at 80 °C for 20 minutes. E. coli DH5α was used for cloning and grown in LB medium at 37 °C with shaking or on LB medium solidified with 1.5 wt% agar supplemented with 100 µg/mL erythromycin.Figure 1

Overview of the pre-culture procedure. In step 1 2.0 wt% glucose was used for the stationary phase pre-culture and 0.5 wt% glucose for the exponential phase pre-culture.

van Tatenhove-Pel R.J., Zwering E., Solopova A., Kuipers O.P, & Bachmann H. (2019). Ampicillin-treated Lactococcus lactis MG1363 populations contain persisters as well as viable but non-culturable cells. Scientific Reports, 9, 9867.

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Plasmid-Based Toxin-Antitoxin Assay

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The experiments were performed as described previously 7 . The assays were performed on Luria-Bertani (LB) medium plates (BD). We used the Escherichia coli BW25113 strain cotransformed with two different plasmid systems for controllable expression of toxins and antitoxins. We used a pair of compatible plasmids: pMG25 for antitoxin expression (high copy number, ColE1 origin of replication (pUC), Amp R , antitoxin expressed under the control of IPTG-inducible P A1/04/03 promoter 43 ] and pBAD33 for toxin expression (medium copy number, p15A origin of replication, Cml R , toxins expressed under the control of arabinose-inducible P BAD promoter 44 ). The cells were grown in liquid LB medium (BD) supplemented with 0.2% glucose (repression conditions), 100 µg ml -1 ampicillin (AppliChem) and 20 µg ml -1 chloramphenicol (AppliChem). Serial dilutions were spotted on solid LB plates supplemented with 0.2% arabinose, as well as 100 µg ml -1 ampicillin (AppliChem) and 20 µg ml -1 chloramphenicol (AppliChem), and bacterial growth was scored after 16-h incubation at 37 °C.

Dominguez-Molina L., Kurata T., Cepauskas A., Echemendia-Blanco D., Zedek S., Talavera-Perez A., Atkinson G.C., Hauryliuk V, & Garcia-Pino A. (2024). Mechanisms of neutralization of toxSAS from toxin-antitoxin modules. Nature chemical biology.

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Subcutaneous Infection of Mice with F. novicida

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Infection of mice with wild-type F. novicida strain U112 was performed at the Biozentrum, University of Basel. All animal experiments were approved (license 2535-26742, Kantonales Veterinäramt Basel-Stadt) and were performed according to local guidelines (Tierschutz-Verordnung, Basel-Stadt) and the Swiss animal protection law (Tierschutz-Gesetz). Bacteria were cultured overnight in brain heart infusion (BHI) medium (supplemented with 100 μg/mL ampicillin [AppliChem] and 0.2% L-cysteine). Bacteria were harvested by centrifugation and washed once with 1× Dulbecco’s Phosphate-Buffered Saline (DPBS). Mice were infected subcutaneously with 5 × 10³ CFU in 50 μL 1× DPBS. Infected mice had access to food and water ad libitum. Mice were sacrificed 48 hr after infection, and spleen and liver were harvested. Colony-forming units (CFUs) were determined from spleen and liver homogenates plated in serial dilutions on Mueller-Hinton agar plates supplemented with 0.1% D-glucose (Millipore), 0.1% fetal calf serum (FCS) (BioConcept), 100 μg/mL ampicillin (AppliChem), and 0.1% L-cysteine. Statistical analysis was performed using GraphPad Prism 6.

Schneider K.S., Groß C.J., Dreier R.F., Saller B.S., Mishra R., Gorka O., Heilig R., Meunier E., Dick M.S., Ćiković T., Sodenkamp J., Médard G., Naumann R., Ruland J., Kuster B., Broz P, & Groß O. (2017). The Inflammasome Drives GSDMD-Independent Secondary Pyroptosis and IL-1 Release in the Absence of Caspase-1 Protease Activity. Cell Reports, 21(13), 3846-3859.

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Cultivation of Candida Species and Bacterial Strains

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C. lusitaniae and C. albicans isolates were grown in YEPD complete medium (1% Bacto peptone; Difco Laboratories, Basel, Switzerland), 0.5% yeast extract (Difco), and 2% glucose (Fluka, Buchs, Switzerland) at 30°C under agitation. The genotypes of all strains constructed are listed in Table S1. Plasmids were propagated in Escherichia coli DH5α on Luria-Bertani (LB) 2% agar plates at 37°C overnight (41 ). LB medium was supplemented with either 100 μg/ml ampicillin (AppliChem) or 34 μg/ml chloramphenicol (Fluka) when necessary.

Kannan A., Asner S.A., Trachsel E., Kelly S., Parker J, & Sanglard D. (2019). Comparative Genomics for the Elucidation of Multidrug Resistance in Candida lusitaniae. mBio, 10(6), e02512-19.

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Bacterial Cloning and Protein Expression

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E. coli strains TOP10 (Invitrogen) and BL21 (DE3) Rosetta (Novagen) were used in cloning and protein expression after transformation by electroporation with the plasmid constructs pT₇-His-sfGFP-mutA and pT₇-His (kindly provided by Prof. Yu Ding, Fudan University, China). For general maintenance and protein expression, E. coli were grown in Luria Broth (LB; 1% Tryptone, 0.5% yeast extract, 171mM NaCl) (Bio Basic INC) with the required antibiotic at 100mg/ml (Ampicillin; Applichem) at 37°C.

Al-Homsi L., Al-Okla S, & Abbady A.Q. (2015). Preparation of Specific Polyclonal Antibody Against the Recombinant Mutacin Produced by sfGFP Fusion Protein Technology. The Open Microbiology Journal, 9, 70-80.

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Bacterial Transformation and Yeast Culture

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E. coli XL1 blue [recA1, endA1, gyrA96, thi-1, hsdR17, supE44, relA1, lac [F´proABlacl q Z DM15 Tn10 (Tetr)] obtained from invitrogen (USA), served as the host strain for bacterial transformation and plasmid isolation. The strain was grown on LB medium (Sigma, USA) supplemented with 75 mg/L ampicillin (Applichem, Germany) or 75 mg/L kanamycin (Roth, Germany) for selection.
In this study, the auxotrophic mutant A. adeninivorans G1212 (aleu2 ALEU2::atrp1—[29 (link)]) and the wild-type A. adeninivorans strain LS3 were used. LS3 was originally isolated from wood hydrolysates in Siberia and deposited as A. adeninivorans SBUG 724 in the strain collection of the Department of Biology of the University of Greifswald [30 (link)]. All strains were grown at 30 °C under non-selective conditions in a complex medium (YEPD) or under selective conditions in yeast minimal medium supplemented with 2 % (w/v) glucose as a carbon source and 43 mM NaNO₃ as a nitrogen source unless stated otherwise (YMM-glucose-NaNO₃) [31 , 32 ].
Agar plates were prepared by adding 1.6 % (w/v) agar to the liquid medium.

Rauter M., Kasprzak J., Becker K., Riechen J., Worch S., Hartmann A., Mascher M., Scholz U., Baronian K., Bode R., Schauer F., Matthias Vorbrodt H, & Kunze G. (2016). Aadh2p: an Arxula adeninivorans alcohol dehydrogenase involved in the first step of the 1-butanol degradation pathway. Microbial Cell Factories, 15, 175.

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Antibody-based Detection of Ribosomal Subunits

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LB medium (5 g yeast extract, 10 g trypton, 5 g NaCl/ l); M9 medium (12.8 g Na₂HPO₄^.7H₂O, 3 g KH₂PO₄, 0.5 g NaCl, 1 g NH₄Cl, 0.4% glucose/ l, 2 mM MgSO₄, 0.1 mM CaCl₂); phosphate buffered saline (PBS) (137 mM NaCl, 2.7 mM KCl, 10 mM Na₂HPO₄, 1.8 mM KH₂PO₄, pH 7.4); S2 and L19 specific antisera, raised in sheep were obtained from Dr. Nierhaus. Horseradish peroxidase (HRP)-conjugated rabbit anti-sheep secondary antibodies were from Jackson ImmunoResearch (CodeNo: 313–035–003; LotNo: 106383). HRP-substrate: for detection a mixture of 1 ml solution A + 100 μl solution B + 1 μl solution C was freshly prepared (solution A: 0.1 mM TRIS (pH8.6), 25 mg Luminol, 100 ml distilled H₂0; solution B: 11 mg p-hydroxycoumaric acid, in 10 ml DMSO; solution C: H₂O₂ (30%). The following antibiotics were used in concentrations as indicated: Ampicillin 100 μg/ml (Applichem-A0839,0100), kanamycin 50 μg/ml (Roth-T832.4), chloramphenicol 7 μg/ml (Sigma-C0378) and erythromycin 100 μg/ml (Sigma-E6367).

Nikolay R., Schloemer R., Schmidt S., Mueller S., Heubach A, & Deuerling E. (2014). Validation of a fluorescence-based screening concept to identify ribosome assembly defects in Escherichia coli. Nucleic Acids Research, 42(12), e100.

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Optimized Heterologous Protein Expression in E. coli

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The gene representing the best hit of the BLAST homology search (vbota) was optimized for E. coli codon usage using an online tool [46 (link)] and synthesized by Eurofins (Eurofins, Luxemburg). The gene was delivered in a pEX-vector. The vector was used to transform E. coli XL1-Blue cells according to the provider's manual. Transformed cells were grown overnight in 3 ml LB (Sigma, Darmstadt, Germany), containing 50 μg/ml ampicillin (Applichem, Darmstadt, Germany). The gene was excised from the vector using NdeI and XhoI (Thermo Scientific, Bremen, Germany) and inserted into the pET21b (+) plasmid (Novagen, Merck, Darmstadt, Germany) behind the inducible T7 promotor. The plasmid was transformed into E. coli XL1-Blue cells. E. coli XL1 transformants were cultivated in 2 ml LB, containing 50 μg/ml ampicillin, overnight, to increase the number of plasmids. After plasmid isolation, positive constructs were used to generate production strains by transforming E. coli BL21 (DE3) cells with them.

Wegner U., Matthes F., von Wirén N., Hajirezaei M.R., Bode R., Kunze G, & Rauter M. (2022). A transaminase with β-activity from Variovorax boronicumulans for the production of enantiopure β-amino acids. Heliyon, 9(1), e12729.

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Bacterial Cultivation and Glycerol Stock Preparation

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Bacterial strains were cultivated in Luria-Bertani (LB) and M9 minimal medium purchased from Becton, Dickinson & Co, Kehl, Germany. Ten milliliters of LB broth supplemented with 200 µg/mL ampicillin was inoculated with a single colony from a freshly streaked plate of Top10 containing BBa_T9002 and incubated for 18 h at 37 °C, shaking at 100 rpm. Each culture was then diluted 1:1000 into 20 mL M9 minimal medium supplemented with 0.2% casamino acids and 1 mM thiamine hydrochloride plus 200 µg/mL ampicillin (AppliChem GmbH, Darmstadt, Germany). The culture was maintained under the same conditions until the optical density measured at 600 nm (OD₆₀₀) reached 0.15 (~5 h). Then 500 µL overnight culture and 500 µL 30% sterile glycerol were mixed together in cryotubes and stored at −80 °C. Before the biosensor assay was conducted, 40 µL bacteria from the glycerol stock vials was cultivated in 20 mL M9 medium plus 200 µg/mL ampicillin until the OD₆₀₀ reached 0.04~0.07 (~4 h).

Qin X., Emich J, & Goycoolea F.M. (2018). Assessment of the Quorum Sensing Inhibition Activity of a Non-Toxic Chitosan in an N-Acyl Homoserine Lactone (AHL)-Based Escherichia coli Biosensor. Biomolecules, 8(3), 87.

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Bacterial Strains and Plasmid Protocols

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The bacterial strains and plasmids used in this study are listed in Table 1. All bacteria were cultured in lysogeny broth (LB; 10 g/l tryptone, 5 g/l yeast extract, 5 g/l NaCl) or on LB agar (15 g/l agar) at 37°C except Serratia plymuthica, which was grown at 30°C. Media were supplemented with the following chemicals (Applichem, Darmstadt, Germany) when appropriate: 5 g/l glucose; 100 μg/ml ampicillin (Ap); 200 μg/ml carbenicillin (Cb); 30 μg/ml chloramphenicol (Cm); 5 μg/ml gentamicin (Gm); 10 μg/ml tetracycline (Tc); 50 μg/ml kanamycin (Km); and 1 mM isopropyl-β-D-thiogalactopyranoside (IPTG). Plasmids pTrc99A and pTrc99A-P_trc-budAB were introduced into the mixed-acid fermenters by electroporation. All oligonucleotides used in this work are listed in Table 2, and were purchased from IDT (Haasrode, Belgium).

Vivijs B., Haberbeck L.U., Baiye Mfortaw Mbong V., Bernaerts K., Geeraerd A.H., Aertsen A, & Michiels C.W. (2015). Formate hydrogen lyase mediates stationary-phase deacidification and increases survival during sugar fermentation in acetoin-producing enterobacteria. Frontiers in Microbiology, 6, 150.

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