All animal experiments were conducted under an animal use protocol approved by the University of California, Davis, Institutional Animal Care and Use Committee (IACUC) or Stanford University, Administrative Panel on Laboratory Animal Care (APLAC). For PET studies and biodistribution, 64Cu-labeled AAVs were evaluated in wild-type 7–9-week-old female C57BL/6 and 9 week-old-female BALB/c mice (The Jackson laboratory, Bar Harbor, ME). For neuraminidase-treated BALB/c mice, neuraminidase (0.12 units/20 μL, Sigma-Aldrich, #N7885) was intranasally administered 3 h before the injection of 64Cu-AAVs. The classical pharmacokinetics and biodistribution studies with qPCR were conducted with wild-type 5–6 week-old female C57BL/6 mice (Charles River).
Balb c mice
Manufactured by Merck Group
Sourced in United States, China
BALB/c mice are an inbred strain of laboratory mice, commonly used in research. They are characterized by a white coat color and are known for their susceptibility to certain diseases and immune responses. BALB/c mice are widely used in various fields of biomedical research, including immunology, oncology, and infectious disease studies.
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Lab products found in correlation
Complete freund s adjuvant, by Merck Group (4 mentions)
Incomplete freund s adjuvant, by Merck Group (2 mentions)
Streptozotocin (stz), by Merck Group (2 mentions)
Neuraminidase, by Merck Group (1 mentions)
Balb c mice, by Jackson ImmunoResearch (1 mentions)
C57bl 6, by Jackson ImmunoResearch (1 mentions)
C57bl 6 mice, by Charles River Laboratories (1 mentions)
Isoflurane anesthesia, by Merck Group (1 mentions)
Dulbecco modified eagle medium (dmem), by Merck Group (1 mentions)
Aluminum hydroxide gel, by Thermo Fisher Scientific (1 mentions)
Female balb c mice, by Orient Bio (1 mentions)
Mouse monoclonal antibody isotyping kit, by Merck Group (1 mentions)
Freund s adjuvant, by Merck Group (1 mentions)
Protein a chromatography, by GE Healthcare (1 mentions)
13 protocols using balb c mice
1
Biodistribution of Cu-AAV in Mice
2
RSV Nucleoprotein Antibody Production
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RSV recombinant nucleoprotein (rNP) (50 µg/100 μL) was mixed with an equal volume of Freund’s complete adjuvant (Sigma-Aldrich, St. Louis, MO, USA) and injected intraperitoneally into six-week-old female BALB/c mice, which were obtained from Orient (Seongnam, Gyeonggi, Korea). Mice were biweekly boosted with RSV rNP (25 μg/100 μL) mixed with an equal volume of Freund’s incomplete adjuvant. The cell fusion technique and indirect ELISA were performed according to previously established protocols [34 (link)]. The isotyping of mAbs was performed with Immuno-Type™ mouse mAb isotyping kit (Sigma-Aldrich) following the manufacturer’s instructions.
3
BALB/c Mice Intranasal Inoculation Assay
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Four-week-old female BALB/c mice (Laboratory Animal Centre of Wuhan Institute of Virology, CAS) were anaesthetized with 250 mg/kg of Averdin (Sigma-Aldrich, St. Louis, USA) before animals were inoculated intranasally with 105 TCID50 of WIV2, WIV3, WIV7 or the DMEM as mock control. Clinical symptoms and body weight were monitored every day up to 21 days. Virus replication and pathogenesis were determined on tissues collected from mice at 1, 3, 5, 7, 10, 14 and 21 days post-infection (dpi). Serum samples were separated from the whole blood by clotting at 37 °C for 1 h and centrifugation at 3,000×g for 10 min.
4
Preclinical Chemotherapy Evaluation in Mice
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For chemotherapy testing, Balb/c mice under isoflurane anesthesia (Sigma-Aldrich Corp.) were challenged with 2 × 105 trypan blue negative P388/S or P388/ADR cells subcutaneously (s.c.) in a volume of 200 μl. Untreated mice were kept in order to determine the lethality of the challenge without chemotherapeutic intervention. Long-term survival was defined as challenged mice that survived the duration of the observation period.
5
Propagation and Titration of MCMV
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The Smith strain of wild type MCMV, which had been passaged in mouse embryonic fibroblasts (MEFs), was provided by Dr. Y. Minamishima (Miyazaki, Japan, [21 (link)]). MEFs were prepared from 12.5-day-old embryos of BALB/c mice (SLC Japan), and were grown in Dulbecco's modified Eagle's essential medium (DMEM; Sigma-Aldrich, Merck, #D5796, Darmstadt, German) containing penicillin (100 units /ml), streptomycin (50 μg/ml), and 10% fetal bovine serum (FBS; Gibco, Thermo Fisher Scientific, #26140079, Waltham, MA). The viral titer was determined by a plaque assay in MEF monolayers as described previously [61 (link)].
6
Immunization Protocol for BALB/c Mice
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Ten female BALB/c mice (16–18 g) (Theodor Bilhariz Research Institute, Giza, Egypt) were immunized with 25 µg polypeptide emulsified with Complete Freund’s adjuvant (Sigma-Aldrich, St. Louis, MI, USA) subcutaneously. The mice were boosted after 21 days with 12.5 µg polypeptide emulsified with InComplete Freund’s adjuvant (Sigma-Aldrich, St. Louis, MI, USA) via the same route then boosted one more time with 12.5 µg polypeptide in PBS intraperitoneally after 2 additional weeks. On day 45, mice were sacrificed, blood was collected by exsanguination of the posterior vena cava, the serum was separated and stored at −20 °C. Mice injected with pyrogen-free saline instead of the polypeptide were used as a negative control.
7
Aggregated MSC1 and MSC1-HI Cell Transplantation
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Prior to transplantation, MSC1 and MSC1-HI cells were cultured and aggregated in non-tissue culture plates using Ham’s F10 media containing supplements for 48 h at 37 °C [17 (link)]. Six million MSC1 or MSC1-HI cells (estimated using a DNA assay [54 (link)]) were transplanted under the left renal subcapsular space of male BALB/c mice rendered diabetic by streptozotocin (270–275 mg/kg of body weight, Sigma Chemical Co., Burlington, MA, USA) injected intraperitoneally 7 days before transplantation, as previously described in Kaur et al., 2014 [16 (link)].
8
Glucosamine Alleviates Atopic Dermatitis in Mice
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This study was carried out in female BALB/c mice (Orient Bio Inc., Seongnam, Korea) aged 8 weeks with AD induced by ovalbumin (OVA; Sigma-Aldrich Korea, Yongin, Korea). Twenty-five BALB/c mice were divided into five groups of five mice: control group without induction of AD (group A), AD group with null treatment by phosphate-buffered saline (PBS) (group B), and AD groups treated with 10 mg, 20 mg, and 40 mg of GlcN (Sigma-Aldrich Korea) administration (groups C, D, and E, respectively). For the AD induction groups (groups B~E), 1.5 ml of OVA and 3 ml of aluminum hydroxide gel (Thermo Fisher Scientific, Waltham, MA, USA) were mixed, and 150 µl of the mixture was intraperitoneally injected into the mice three times a week for three weeks. After a week of OVA injection, mice were epicutaneously sensitized with OVA patches. The patches were made by dripping 50 µl of OVA (1 mg/ml) in a gauze (1×1 cm) which was placed on the back skin three times per week for two weeks. Group A was injected with PBS instead of OVA. This experiment was conducted in specific-pathogen-free environment and the mice received an OVA-free diet. This study was reviewed and approved by the Institutional Animal Care and Use Committee of Inha University (INHA 181120-601).
9
Diabetic Foot Ulcer Mouse Model
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We established the mice model of DFU according to the previous study of Li [22 (link)]. In general, male BALB/c mice (Shanghai Laboratory Animal Center of Chinese Academy of Sciences, China) with 5 weeks age and 19.39 ± 1.56 g weight were treated with 0.45% streptozotocin (Sigma, USA, 45 mg/kg) and feed with high-fat diet. We choosed these mice with the blood glucose level at 16.7 mmol/L to establish DFU model. There were 62 mice with DM. Then, we used the sterile punch to make wounds. Finally, exosomes derived from mmu_circ_0001052-modified ADSCs (exosomes + mmu_circ_0001052 group) or exosomes from vector-modified ADSCs (exosome + vector group) were subcutaneously injected. 25 μL of exosomes (200 μg in 100 μL PBS) were subcutaneously injected at 4 sites around the wound. The control group was not treated with exosomes. Wound tissues were collected at 3, 7 and 14 days after wounding. This study, following the Nation Institutes of Health Guide for the Laboratory Animals Care and Use, was carried out with the approval of hospital ethics committee, and Ethical approval No.:Med-Eth-Re [2021] 192.
10
Immunization and Antibody Titration Protocol
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Purified rPDHA and rPDHB proteins, and M. gallisepticum whole cells (inactivated with 0.4% formalin for 16 h at 37°C) were emulsified with an equal volume of Freund’s complete adjuvant (Sigma) and used to immunize 6-week-old female BALB/c mice (SLAC, Shanghai, China) via multipoint subcutaneous injection respectively (100 μg purified protein or 1010 colony forming units [CFUs] M. gallisepticum whole cells per mouse). After the first immunization, two boosters were given at 2-week intervals. Two non-immunized mice were used as negative controls. Tail vein blood from immunized and non-immunized mice was collected and the titres of polyclonal antisera measured by indirect enzyme-linked immunosorbent assay (iELISA) as previously described [22 (link)], with 96-well plates coated with coating buffer (16 mM Na2CO3, 34 mM NaHCO3, pH 9.6) containing purified protein (0.5 μg per well) or M. gallisepticum total protein prepared by sonic disruption of M. gallisepticum bacteria (0.5 μg per well) overnight at 4°C. When titres significantly increased, blood samples were collected from the infraorbital sinuses of mice and serum samples were separated and stored at −20°C.
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