Nisin Z (2.5% w/w, N5764), Pepsin (P0525000), MTT (M2003), and Cytotoxicity Detection Kit PLUS (LDH) were obtained from Sigma Aldrich (Milan, Italy). FITC Annexin-V apoptosis kit I (BD Pharmigen™, BD Biosciences, and San Jose, CA, USA, 556547) and fetal bovine serum (FBS) were purchased of Cegrogen-Biotech (Germany, A0100-3010). Pen-Strep (10,000 Unit/mL penicillin and 10,000 unit/mL streptomycin) and 0.25% trypsin-EDTA were purchased from Bioidea (Tehran, Iran). RPMI was prepared from GIBCO Laboratories (Grand Island, NY, USA, 11530586). Coumarin-6 and Oxaliplatin were kindly gifted by Professor Valizadeh’s laboratory (TUOMS, Tabriz, Iran). All other chemicals were of analytical grade.
Nisin z
Manufactured by Merck Group
Sourced in Italy
Nisin Z is a purified natural antimicrobial peptide produced by certain strains of Lactococcus lactis. It exhibits broad-spectrum antimicrobial activity against Gram-positive bacteria, including food-borne pathogens and spoilage organisms.
Automatically generated - may contain errors
Lab products found in correlation
Pepsin, by Merck Group (2 mentions)
Cytotoxicity detection kitplus ldh, by Merck Group (1 mentions)
Fitc annexin 5 apoptosis kit 1, by BD (1 mentions)
Pen strep, by Bioidea (1 mentions)
Trypsin edta, by Bioidea (1 mentions)
Roswell park memorial institute (rpmi), by Thermo Fisher Scientific (1 mentions)
Methyl thiazolyl tetrazolium (mtt), by Merck Group (1 mentions)
96 well plate, by Sarstedt (1 mentions)
Infinite m100, by Tecan (1 mentions)
4 protocols using nisin z
1
Cytotoxicity and Apoptosis Evaluation
2
Antimicrobial Activity of Nisin Z and ILs
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Nisin Z (purity ≥98%)
(Figure 1 a) was purchased
from Sigma-Aldrich. Luria–Bertani broth (LBB), E. coli (MTCC 40), and S. aureus (MTCC 87) cultures were obtained from the microbial type culture
collection and gene bank (MTCC), CSIR-Institute of microbial technology,
Chandigarh, India. The ILs used in present study were 1-butyl-1-methylpyrrolidin-1-ium
bis(trifluoromethylsulfonyl)imide [Pyr C4]NTF2, 1-hexyl-1-methylpyrrolidin-1-ium bis(trifluoromethylsulfonyl)imide
[Pyr C6]NTF2, 1-methyl-1-octylpyrrolidin-1-ium
bis(trifluoromethylsulfonyl)imide [Pyr C8]NTF2, 1-decyl-1-methylpyrrolidin-1-ium bis(trifluoromethylsulfonyl)imide
[Pyr C10]NTF2, and 1-dodecyl-1-methylpyrrolidin-1-ium
bis(trifluoromethylsulfonyl)imide [Pyr C12] NTF2 (Figure 1 b), which
were synthesized in our laboratory.12 (link) Ethanol,
glycerol, and autoclaved water were used to study the antibacterial
activity, while Milli-Q water was used for the spectroscopic and tensiometry
experiments.
(
from Sigma-Aldrich. Luria–Bertani broth (LBB), E. coli (MTCC 40), and S. aureus (MTCC 87) cultures were obtained from the microbial type culture
collection and gene bank (MTCC), CSIR-Institute of microbial technology,
Chandigarh, India. The ILs used in present study were 1-butyl-1-methylpyrrolidin-1-ium
bis(trifluoromethylsulfonyl)imide [Pyr C4]NTF2, 1-hexyl-1-methylpyrrolidin-1-ium bis(trifluoromethylsulfonyl)imide
[Pyr C6]NTF2, 1-methyl-1-octylpyrrolidin-1-ium
bis(trifluoromethylsulfonyl)imide [Pyr C8]NTF2, 1-decyl-1-methylpyrrolidin-1-ium bis(trifluoromethylsulfonyl)imide
[Pyr C10]NTF2, and 1-dodecyl-1-methylpyrrolidin-1-ium
bis(trifluoromethylsulfonyl)imide [Pyr C12] NTF2 (
were synthesized in our laboratory.12 (link) Ethanol,
glycerol, and autoclaved water were used to study the antibacterial
activity, while Milli-Q water was used for the spectroscopic and tensiometry
experiments.
3
Antimicrobial Activity of Nisin Z and Pepsin
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Nisin Z (2.5% w/w) and pepsin were obtained from Sigma Aldrich (Milan, Italy). β-CD was purchased from Cyclolab (Budapest, Hungary). E. coli (ATCC 25992) and S. aureus (ATCC 25923) were prepared from a Persian collection of bacteria in the Iranian Research Organization for Science and Technology (Tehran, Iran). All other reagents used were of analytical grade.
4
Measuring Antimicrobial Peptide Resistance
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To assess resistance to AMPs, a growth inhibition assays were performed in micro titer plate format as described previously (Wiegand et al., 2008) (link) with minor modifications. Serial 1:2 dilutions of a standard solution of nisin Z (Sigma-Aldrich) were prepared in 2xTY and 100 µl were distributed in 96-well plates (Sarstedt, Nümbrecht, Germany). Bacteria of the test strains were inoculated from agar plates and cultivated overnight in 2xTY medium containing kanamycin. IPTG (0.1 mM) and/or ATC (0.25 µg/ml) were added to induce gene expression where appropriate. o/N cultures were diluted in fresh medium containing the inducers and 100 µl were added to each well of the assay plate to a final optical density at 600 nm (OD600) of 0.05 in the assay. Assay plates were incubated with agitation at 30 °C. Growth was determined after 24 h by measuring OD600 in an Infinite M100 plate reader (Tecan, Männedorf, Suisse). The minimal inhibitory concentration (MIC) was calculated based on the lowest dilution of the nisin standard.
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