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47 protocols using balb c mice

1

Mouse Infection Model for H5N8 and H5N1

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Six-week-old female BALB/c mice (Samtako, Seoul, Korea) were used for all infection experiments. Ten mice per group were inoculated intranasally with 102–107 EID50/50 mL of MDk/W452(H5N8) and EM/W149(H5N1). Body weights and survival patterns were recorded daily for 14 days. For virological and pathological examinations, groups of mice were inoculated intranasally with 105 EID50/50 mL of virus, and six mice per group were euthanised at 1, 3, 5, 7 and 9 dpi to examine the growth kinetics of the virus in mouse lungs.
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2

Antibody Response of BALB/c Mice

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Six-week-old female BALB/c mice were purchased from Samtako (Osan, Korea). The experiments were performed in accordance with the guidelines for the care and use of laboratory animals under the approval of animal ethics committee at Seoul National University (SNU-130415–1). The mice, maintained under standard pathogen-free conditions, were provided with free access to food and water during the experiments. Mice (n = 5) were immunized subcutaneously with 10 μg of purified proteins three times at 2-week intervals. Complete Freund’s adjuvant (CFA) and incomplete Freund’s adjuvant (IFA) were mixed with antigens for the priming and boosting of animals. Sera were collected from the mice at 0, 14, 28 and 42 days after first immunization for the antibody detection and serum neutralization assay.
The induction of antigen specific serum IgG level was measured by ELISA. Plates were coated and blocked using the same method described in the indirect ELISA section. After blocking, 2 fold serial diluted mouse sera were added to the wells and incubated for 1 h at room temperature. HRP conjugated goat anti-mouse IgG secondary antibody diluted at 1:5000 was added and incubated for 1 h at room temperature. Other assay procedures are same as described in the indirect ELISA section.
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3

PM2.5-Induced Lung Injury: Evaluating Weed Extract

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BALB/c mice (male, 6 weeks) were obtained from Samtako (Osan, Korea), and mice were randomly assigned to four per cage (a 12 h light/dark cycle, 55% humidity, and 22 ± 2 °C). Each group (n = 14) was divided into 6 groups, and each consisted of a normal control (NC) group, a PM2.5-exposed group (negative control group; PM2.5), and WEE groups (50, 100, and 200 mg/kg of body weight, respectively; WEE 50, WEE 100, and WEE 200). The WEE was dissolved into drinking water and was orally fed using a stomach tube once a day for 12 weeks. WHO Air quality guidelines for fine particulate matter recommend expositions to PM2.5 < 25 μg/m3 24 h mean in human, and the annual PM2.5 concentration in the most polluted cities in the world is around 170 μg/m3 [56 ,57 ]. The mice were exposed to PM2.5 at a 500 μg/m3 concentration in the whole-body exposure chamber (5 h/day and 5 days/week for 12 weeks).
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4

PCV2 Vaccine Efficacy in Mice

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The experimental design is displayed in Table 2. Six-week-old female BALB/c mice were purchased from Samtako, Korea. The mice were SPF grade (specific pathogen-free) and were randomly allocated into three groups of six mice and housed in filter-top cages in isolation rooms under controlled temperature (20-25 o C), humidity (60-80%), and fed sterilized food and water. Groups 1 and 2 were intramuscularly vaccinated with 10 9 PFU of Bac-Rep-Cap or 10 9 PFU of Bac-Cap with Freund's complete adjuvant (Sigma-Aldrich, USA) at a ratio of 50:50 (w/w) according to the manufacturer's instructions. Group 3, the negative control group, was immunized with 100 μl of PBS. At 21 and 42 days after the primary immunization, the serum samples were collected for detection of antibody to PCV2 by ELISA and analyzed for the presence of PCV2-specific neutralizing antibodies. All experiments were done with the approval of the Institutional Animal Care and Use Committee of Catholic University of Pusan (CUP 2018-2).
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5

Immunization Study of Female BALB/c Mice

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Six-week-old female naïve BALB/c mice (17–19 g) were purchased from Samtako (Korea). The mice were maintained under standard pathogen-free conditions on a 12:12 h light/dark cycle (5 mice per cage), monitored three times weekly and provided with free access to food and water during experimentation. All mice were divided randomly into different groups before immunization and adapted for 1 week before any treatments. Animal experiments were performed in accordance with the guidelines for the care and use of laboratory animals under the approval of the animal ethics committee at Seoul National University (No. SNU-161114-4). After the final serum collection of each experiment, the mice were euthanized with CO2 inhalation.
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6

Murine Macrophage Isolation and LPS Response

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Seven-week-old male BALB/c mice were obtained from Samtako (Osan, Korea) and housed in a temperature- and humidity-controlled pathogen-free animal facility with a 12 h light-dark cycle. The animal protocol was approved by the Institutional Animal Care and Use Committee at Kyung Hee University (KHUASP(SE)-15-012), and the mice were cared for according to the US National Research Council for the Care and Use of Laboratory Animals (1996) specifications. All animals underwent 1 week of adjustment prior to experiments and were randomly allocated to the experimental groups. RVS extract dissolved in water (20 mg/mL or 100 mg/mL) was given orally to mice at doses of 200 mg/kg or 1000 mg/kg body weight once daily for 10 days. Control mice were given an equal volume of water. Mice were assigned for two experiments: peritoneal macrophage isolation (N = 6) and intraperitoneal injection of LPS (N = 12). These procedures are given below.
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7

Immunization Protocols for BALB/c Mice

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Female 6 weeks old BALB/c mice were purchased (Samtako, Korea) and grouped according to the experimental plan. Mice were housed in ventilated cages and maintained with pelleted feed and tap water ad libitum. Mice immunization was carried out with different treatment combinations as displayed in Table 2. Blood samples were collected from the retro-orbital sinus at different time points as described in the vaccination schedule (Fig. 2A). The collected blood samples were then centrifuged at 12000 rpm for 10 min at 4 °C and the serum was separated and stored at -20 °C until use. At day 42 following the first immunization, five mice from each group were sacrificed to collect splenocytes and PBMC.
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8

BALB/c Mice for Pathogen Research

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Female-specific pathogen-free 6-week-old BALB/c mice were purchased from SAMTAKO (Osan, Korea). The mice were maintained under standard conditions (on a 12 h night/day cycle, at a humidity of 55 ± 5 % and temperature of 22 ± 2 °C) and fed ad libitum. All experiments were conducted according to a protocol approved by the Chonnam National University Institutional Animal Care and Use Committee. (CNU IACUC-YBR-2016-19, Gwangju, Korea)
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9

LPS-Induced Lung Inflammation Timepoints

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Using the same methods, two animal studies were conducted at different times. Eighty female BALB/c mice were purchased from Samtako (Osan, Korea) and based on the elapsed time after 20 µg/20 µL/head LPS intranasal instillation, they were divided into 5 groups: (1) 0 hr, (2) 3 hr, (3) 24 hr, (4) 48 hr, and (5) 72 hr (Figure 1).
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10

Murine Bordetella bronchiseptica Model

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C57BL/6 and Balb/c mice were purchased from Samtako (Korea) and housed at the animal facility of Jeju National University. In this study, 7- to 12-week-old female mice were used for experiments. Animal experiments in this study were performed in accordance with the Institutional Guidelines for Animal Use and Care of Jeju National University (2021-0015). DOX and VC were purchased from Sigma-Aldrich (St. Louis, MO, USA) and dissolved in dimethyl sulfoxide and phosphate buffer solution (PBS), respectively. Bordetella bronchiseptica in PBS was sonicated under conditions of amplitude 60% for 3 cycles of 30 sec with a 30 sec interval using a Q125 sonicator (Qsonica, Newtown, CT, USA), then the amount of protein was measured by the Bradford assay and the proteinsBb was stored in a −20 °C freezer.
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