Cronobacter sakazakii strains (strain no. ATCC29544, ATCC12868, ATCC29004, and ATCCBAA-894) and human brain microvascular endothelial cells (HBMEC) were purchased from the American Type Culture Collection (ATCC). Bacterial strains were grown at 37°C in brain heart infusion (BHI) broth (Landbridge, Beijing, China) overnight. HBMEC were grown in DMEM (Dulbecco’s modified eagle medium, Gibco, Grand Island, NY, United States) with 10% FBS (Biological Industries, Kibbutz Beit Haemek, Israel) at 37°C and 5% CO2 for the indicated period of time in different experiments.
Atcc 29004
Manufactured by American Type Culture Collection
Sourced in United States
ATCC 29004 is a microplate designed for cell culture applications. It is made of polystyrene and has 96 wells with a flat bottom. The microplate is transparent, allowing for optical measurements and observations.
Automatically generated - may contain errors
Lab products found in correlation
Atcc 29544, by American Type Culture Collection (4 mentions)
Atcc12868, by American Type Culture Collection (2 mentions)
Atccbaa 894, by American Type Culture Collection (2 mentions)
Dulbecco s modified eagle medium, by Thermo Fisher Scientific (1 mentions)
Hbmec, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Sartorius (1 mentions)
Hbmec, by American Type Culture Collection (1 mentions)
Agno3, by Sinopharm (1 mentions)
Glycerol, by Sangon (1 mentions)
6 protocols using atcc 29004
1
Cronobacter sakazakii Infection of HBMEC
2
Coenzyme Q10 Inhibits Cronobacter Biofilm
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CoQ 0 (HPLC ≥99%, CAS 605-94-7) was obtained from J&K Scienti c Co., Ltd (Beijing, China) and dissolved in dimethyl sulfoxide (DMSO) for use in all experiments. The nal concentration of DMSO in all sample solutions was 0.1% (v/v), which has no apparent effect on the growth of C. sakazakii. All other chemicals were of analytical grade and were unaltered.
Bacterial strains and culture conditions C. sakazakii strains ATCC 29004 and ATCC 29544 were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA). Strain ATCC 29004, which is a relatively strong bio lm producer, was used in the bio lm assay. Prior to each assay, bacteria were inoculated onto tryptic soy agar (TSA) medium and incubated at 37°C for 12 h. To obtain fresh overnight cultures, a single colony was inoculated into 30 mL of tryptic soy broth (TSB) medium and incubated with shaking at 130 rpm for 12 h at 37°C. Following incubation, cultures were centrifuged (4°C, 8000 × g, 5 min), washed three times with sterile phosphate-buffered saline (PBS), and diluted in TSB medium to an optical density at 600 nm (OD 600 ) of 0.5 (approximately 4 × 10 8 colony-forming units (CFU)/mL).
Bacterial strains and culture conditions C. sakazakii strains ATCC 29004 and ATCC 29544 were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA). Strain ATCC 29004, which is a relatively strong bio lm producer, was used in the bio lm assay. Prior to each assay, bacteria were inoculated onto tryptic soy agar (TSA) medium and incubated at 37°C for 12 h. To obtain fresh overnight cultures, a single colony was inoculated into 30 mL of tryptic soy broth (TSB) medium and incubated with shaking at 130 rpm for 12 h at 37°C. Following incubation, cultures were centrifuged (4°C, 8000 × g, 5 min), washed three times with sterile phosphate-buffered saline (PBS), and diluted in TSB medium to an optical density at 600 nm (OD 600 ) of 0.5 (approximately 4 × 10 8 colony-forming units (CFU)/mL).
3
Activation and Standardization of C. sakazakii
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C. sakazakii strain ATCC 29004 was purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA) and was preserved in tryptone soya broth (TSB, Land Bridge Technology, Beijing, China) containing 20% (v/v) glycerol at −80°C. For activation, the stock culture was inoculated onto trypticase soy agar (TSA, Land Bridge Technology, Beijing, China) and cultured at 37°C overnight. Then a single colony was inoculated into TSB and the culture was incubated at 37°C, with shaking at 180 rpm, for 8 h. After activation, the cells from the culture were obtained by centrifugation (5000 × g, 4°C, 15 min). Precipitates were washed twice with phosphate-buffered saline (PBS, Land Bridge Technology, Beijing, China), and finally resuspended and adjusted to an OD600nm = 0.5, with a cell concentration of 108 CFU/mL. Finally, the bacterial suspensions were diluted to a concentration of 107 CFU/mL prior to mature biofilm formation.
4
Synthesis and Characterization of Silver Nanoparticles
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The silver to produce AgNP was from silver nitrate (AgNO 3 , Sinopharm Chemical Reagent Co. Ltd., Shanghai, China), using citrate sodium as the reducing agent (Fuchen Chemical Reagent Factory, Tianjin, China); glycerol and PVP (Sangon Biotech Co. Ltd., Shanghai, China) were used as stabilizers. All chemicals were of analytical grade. Four C. sakazakii strains: ATCC 29544 T , ATCC BAA894, ATCC 29004, and ATCC 12868, were purchased from American Type Culture Collection (ATCC, Manassas, VA) and preserved in our laboratory at -80°C. Each strain was inoculated into 20 mL of Luria-Bertani (LB) medium and incubated for 12 h at 37°C with shaking at 160 rpm.
5
Cultivation and Standardization of C. sakazakii
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C. sakazakii strains ATCC 29004 and ATCC 29544 were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA). Strain ATCC 29004, which is a relatively strong biofilm producer, was used in the biofilm assay. Prior to each assay, bacteria were inoculated onto tryptic soy agar (TSA) medium and incubated at 37°C for 12 h. To obtain fresh overnight cultures, a single colony was inoculated into 30 mL of tryptic soy broth (TSB) medium and incubated with shaking at 130 rpm for 12 h at 37°C. Following incubation, cultures were centrifuged (4°C, 8000 × g, 5 min), washed three times with sterile phosphate-buffered saline (PBS), and diluted in TSB medium to an optical density at 600 nm (OD 600 ) of 0.5 (approximately 4 × 10 8 colony-forming units (CFU)/mL).
6
Characterization of Cronobacter sakazakii Strains
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C. sakazakii strains ATCC 29544 and ATCC 29004 were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). Three other C. sakazakii strains (CS 1, CS 2, CS 3) were taken from our laboratory strain collection, which were originally isolated from infant formula and infant rice cereal collected from supermarkets in China. All isolates were used in minimum inhibitory concentration assay and only C. sakazakii ATCC 29544 was used for antimicrobial mechanism analysis. All strains were stored in tryptone soya broth (TSB) with 20% glycerol (v/v) at -80°C. Before each experiment, stock cultures were streaked on tryptone soya agar (TSA) and grew at 37°C for 18 h. Then a loopful of each strain was inoculated into 30 mL TSB and incubated for 18 h at 37°C.
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