A total of three male C57BL/6 mice (age, 4–6 weeks; weight, 10–16 g; homozygote PDK-1flox/flox) were designed and provided by GemPharmatech Co., Ltd. The upstream and downstream sequences of the PDK-1 gene were inserted into the loxP site. The results of the identification were provided by GemPharmatech Co., Ltd. and validated in the present study (shown in Fig. S1 ). A total of three male C57BL/6 mice (age, 4–6 weeks old; weight, 10–16 g; wild-type, PDK-1+/+) were also purchased from GemPharmatech Co., Ltd. All animals had the same genetic background. All mice were raised and maintained in the laboratory animal center of Guangxi Medical University (Nanning, China). Mice were individually housed at a controlled temperature (22–26°C) and humidity (50-60%) in ventilated cages with a 12-h light/dark cycle and free access to standard chow and fresh water. Experiments on all animals were conducted according to the 2013 ARRIVE and AVMA guidelines for euthanasia. Briefly, mice were anesthetized via inhalation of 2% sevoflurane and were placed in a container; CO2 was then injected into the container at a rate that replaced 25% of container volume per minute. When it was confirmed that the heartbeats of the mice had stopped and they were no longer breathing, the CO2 was turned off. Finally, the mice were observed for 2 min to confirm death. The mice were sacrificed in November 2019.
C57bl 6 mice
Manufactured by GemPharmatech
Sourced in China
C57BL/6 mice are a common inbred mouse strain widely used in biomedical research. They are a well-characterized model organism with a stable and consistent genetic background.
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Lab products found in correlation
One step mouse genotyping kit, by Vazyme (2 mentions)
App ps1 jax, by Jackson ImmunoResearch (2 mentions)
Lipopolysaccharides (lps), by Merck Group (2 mentions)
D12492, by Research Diets (2 mentions)
Trizol, by Thermo Fisher Scientific (1 mentions)
C57bl 6 mice, by Japan SLC (1 mentions)
Be0089, by BioXCell (1 mentions)
Be0146, by BioXCell (1 mentions)
Balb c nude mice, by GemPharmatech (1 mentions)
Ob ob mice, by GemPharmatech (1 mentions)
Methylprednisolone, by Pfizer (1 mentions)
Nude mice, by GemPharmatech (1 mentions)
Balb c, by GemPharmatech (1 mentions)
Isoproterenol hydrochloride, by Merck Group (1 mentions)
Db db mice, by GemPharmatech (1 mentions)
Apoe mice, by GemPharmatech (1 mentions)
Hepa1 6 cells, by American Type Culture Collection (1 mentions)
C57bl 6, by Shanghai Model Organisms (1 mentions)
Cerulean, by Merck Group (1 mentions)
I5627, by Merck Group (1 mentions)
115 protocols using c57bl 6 mice
1
Generation and Validation of PDK-1 Knockout Mice
2
Murine Model of Echinococcus multilocularis Infection
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Eight- to ten-week-old C57BL/6 mice and IDO1-deficient (IDO1-/-) C57BL/6 mice were purchased from the Jiangsu GemPharmatech Co. Ltd., and were bred in a specific pathogen-free environment with a 12-h light/dark cycle supplemented with rodent chow and water ad libitum.
E. multilocularis protoscoleces (PSCs) were obtained from intraperitoneal lesions maintained in BALB/c mice under aseptic conditions, followed by three rounds of clean-up with phosphate buffered saline (PBS, pH=7.2, containing 1000 mg/mL penicillin and 1000 U/mL streptomycin) (36 (link)). PSCs with over 95% vitality determined by eosin exclusion were counted using hemocytometer (37 (link)), and 1000 PSCs were injected intraperitoneally per mice as previously described (38 (link)). To determine the parasitic burden using wet-weighing method, mice were sacrificed in month 1, 2 or 3 post-infection, and parasitic tissues were dissected from the liver and the peritoneal cavity.
E. multilocularis protoscoleces (PSCs) were obtained from intraperitoneal lesions maintained in BALB/c mice under aseptic conditions, followed by three rounds of clean-up with phosphate buffered saline (PBS, pH=7.2, containing 1000 mg/mL penicillin and 1000 U/mL streptomycin) (36 (link)). PSCs with over 95% vitality determined by eosin exclusion were counted using hemocytometer (37 (link)), and 1000 PSCs were injected intraperitoneally per mice as previously described (38 (link)). To determine the parasitic burden using wet-weighing method, mice were sacrificed in month 1, 2 or 3 post-infection, and parasitic tissues were dissected from the liver and the peritoneal cavity.
3
Trem2 Knockout Mouse Model of Schistosomiasis
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B6/JGpt‐Trem2em1Cd3332in1/Gpt knockout mice (Trem2−/− mice) were established by Gempharmatech Co., Ltd (Jiangsu, China) and wild‐type C57BL/6 mice were purchased from Laboratory Animal Centre of Nantong University. All mice were raised in the barrier environment of the Laboratory Animal Centre at Nantong University. Snails infected with S.japonicum were provided by National Institute of Parasitic Diseases, Chinese Centre for Disease Control and Prevention. To establish the S. japonicum infection models, mice were infected with cercariae S. japonicum (15 ± 2) via abdominal skin exposure. The mice were randomly divided into various groups (n≧3) and sacrificed at the indicated time. The liver tissues harvested for RNA extraction were preserved in Trizol (Thermo Fisher Scientific) at −80°C. The liver tissues for western blotting were harvested in a tube directly and preserved at −80°C. The liver tissues for immunofluorescence staining and immunohistochemistry were harvested and immersed in formalin solution at 4°C. Peritoneal macrophages were obtained from mice as Xu et al. reported.12
4
C57BL/6 Mouse Acclimation Protocol
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C57BL/6 mice were obtained from Gempharmatech Co., Ltd (Nanjing, China). The mice were allowed to acclimate to the living environment for 7 days prior to the start of all experiments.
5
Animal Models for Lung Fibrosis and Injury
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Mice. For the bleomycin-induced lung fibrosis model and the LPS-induced acute lung injury model, C57BL/6 mice (male, 8 weeks old, ~23-26 g) were purchased from Jiangsu Gempharmatech. Animals were acclimatized for 1 week before the experiment. All in vivo experimental procedures were approved by the IACUC. All euthanasia was performed using CO 2 inhalation, and all efforts were made to minimize animal suffering.
For the UUO study, 7-week-old female C57BL/6 mice were obtained from Japan SLC. Animals were housed and fed a normal diet (CE-2, CLEA Japan) under controlled conditions. All animals used in the study were housed and cared for following the Japanese Pharmacological Society Guidelines for Animal Use.
For the UUO study, 7-week-old female C57BL/6 mice were obtained from Japan SLC. Animals were housed and fed a normal diet (CE-2, CLEA Japan) under controlled conditions. All animals used in the study were housed and cared for following the Japanese Pharmacological Society Guidelines for Animal Use.
6
Anesthesia and Euthanasia in Murine Study
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We used a total of 120 mice in our study. Each experimental and control group comprised 6 mice, based on our previous experience. Detailed information regarding the allocation of mice to specific experimental conditions is provided in the Materials and methods section. All female C57BL/6 mice (7–8 weeks old), purchased from GemPharmatech Co., Ltd. (China), were housed under specific pathogen-free (SPF) conditions. All mice were included in the experiment only after being confirmed to be free of any fundus or systemic diseases. Animal care and experimentation were conducted in accordance with the Association for Research in Vision and Ophthalmology (ARVO) Statement. In this study, tribromoethanol was used for anesthetizing mice, and cervical dislocation was employed for euthanizing the mice. All animal procedures were approved by the Animal Care and Use Committee of Tianjin Medical University Eye Hospital (TMUEC).
7
Murine Model of Pathogen-Free Research
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All animal experiments were performed in accordance with international guidelines and approved by the Institutional Animal Care and Use Committee of the PLA General Hospital. Male C57BL/6 mice aged 6–8 weeks were purchased from GemPharmatech Co., Ltd. Mice were housed in a pathogen-free, constant temperature environment with a 12 h light/dark cycle and allowed to acclimatize for a week in the animal facility before the operation.
8
Murine Breast Cancer Xenograft Model
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Six-week-old female C57BL/6 mice were purchased from GemPharmatech (Jiangsu, China). The mice received tumor cell inoculation after 1 week of acclimation. The Institutional Animal Care and Use Committee authorized all procedures for the use and care of animals. The Py230 and Py8119 syngeneic murine breast cancer cells were trypsinized, washed twice in phosphate-buffered saline (PBS), counted, and resuspended in 1:1 Matrigel:PBS before being injected (5 × 106 cells/mouse) into the fourth mammary fat pad on the right side of the mice. The tumor volume was calculated using the formula V = (W2 × L)/2 from caliper measurements, where V is the tumor volume, W is the tumor width, and L is the tumor length.
9
Transgenic Mouse Models for Cell Fate Tracing
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The LmnaG609G/G609G, Lmnaf/f;TC and Tie2–cre mice were as previously described37 (link). ROSAmT/mG mice were provided by Dr Jian Chen (Suchow University, China). C57BL/6 mice, H11-mG/mR mice and Cdh5-cre/ERT mice were purchased from GemPharmatech. All mice were housed and handled in accordance with protocols approved by the Committee on the Use of Live Animals in Teaching and Research of Shenzhen University.
10
Murine Paraspinal Muscle Development
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C57BL/6 mice were obtained from GemPharmatech (Jiangsu, China) and housed in a temperature/light-controlled environment with access to standard mouse chow and water ad libitum. Embryonic day 16.5 (E16.5), E17.5, and postnatal day 0 (P0) embryos were microdissected to obtain the dorsal paraspinal muscles. The female mice were dissected at month 1 (M1), M6, M12, and M24 to obtain the paraspinal muscles.9 (link) The collected tissues were washed with 1× phosphate-buffered saline (PBS), and then snap-frozen and stored in liquid nitrogen prior to nuclear extraction. The nuclei were separated by mechanical extraction.10
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