Toxoplasma gondii RH strain and ME49 strain were maintained according to the methods described previously [18 (link)–20 (link)]. T. gondii RH stain was used for RNA extraction, and T. gondii ME49 was used to infect mice and to collect sera. Spodoptera frugiperda Sf9 cells were maintained in suspension in serum-free SF900 II medium (Invitrogen, Carlsbad, California, USA) at 27°C in spinner flasks at 130 to 140 rpm. S. frugiperda Sf9 cells were used for production of recommended baculovirus (rBV) and virus-like particles. Horseradish peroxidase (HRP)-conjugated goat anti-mouse immunoglobulin G (IgG) was purchased from Southern Biotech (Birmingham, Alabama, USA).
Horseradish peroxidase hrp conjugated goat anti mouse immunoglobulin g igg
Manufactured by Southern Biotech
Sourced in United States
Horseradish peroxidase (HRP)-conjugated goat anti-mouse immunoglobulin G (IgG) is a laboratory reagent used in various immunoassays and detection methods. It consists of goat-derived IgG antibodies that are covalently linked to the enzyme horseradish peroxidase. This conjugate can be used to detect the presence of mouse IgG in biological samples.
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4 protocols using horseradish peroxidase hrp conjugated goat anti mouse immunoglobulin g igg
1
Maintenance and Infection Protocols for Toxoplasma gondii
2
Serological Evaluation of T. gondii VLP
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Female inbred BABL/c (aged 8 weeks; Nara Biotech, Kyonggi-do, Korea) were used. All animal experiments and husbandry involved in these studies were conducted under the guidelines of the Kyung Hee University IACUC. Mice were infected with T. gondii (ME49), and the mouse sera were collected at week 4 and stored at −20°C until used. VLPs containing T. gondii ROP4 protein was identified by western blot. Monoclonal mouse anti-M1 antibody was used to confirm influenza M1 protein content. The levels of IgG antibody was determined by ELISA. Flat-immunoplates (96-well) were coated with 100 μl of VLPs at a concentration of 4 μg/ml in 0.05 M carbonate bicarbonate buffer (pH 9.6) per well and incubated overnight at 4°C. The mouse serum samples were serially diluted in PBST (100 μl/well) and incubated in the plates for 1.5 hr at 37°C. Horseradish peroxidase (HRP)-conjugated goat anti-mouse immunoglobulin G (IgG) was purchased from Southern Biotech.
3
Murine Malaria Infection Model
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Six-week-old female BALB/c mice were obtained from NARA Biotech (Seoul, Korea). The Plasmodium berghei ANKA strain 2.34 was maintained in mice by serial intraperitoneal passage. Spodoptera frugiperda Sf9 cells were used for the production of recombinant baculovirus (rBV) and VLPs. They were maintained in serum-free SF900 II medium (Invitrogen, Carlsbad, CA, USA) in spinner flasks at 27 °C and 130–140 rpm. P. berghei-infected sera from mice were collected through retro-orbital plexus puncture. Horseradish peroxidase (HRP)-conjugated goat anti-mouse immunoglobulin G (IgG) was purchased from Southern Biotech (Birmingham, AL, USA). Monoclonal mouse anti-M1 antibody was purchased from Abcam (Cambridge, UK).
4
Maintaining Plasmodium berghei Infection and Vaccine Immunization in Mice
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To maintain the Plasmodium berghei ANKA strain and immunization vaccines, female seven-week-old BALB/c mice were purchased from NARA Biotech (Seoul, Korea). Mice were housed in an institute-approved facility with a 12 h day and night cycle, with easy access to food and water. Nesting material and enrichment were also provided to prevent the development of abnormal behaviors.The P. berghei ANKA strain was used for challenge infection and antigen preparation. CV1 cells (CCL-70) and Vero cells (CCL-81) were purchased from the American Type Culture Collection (Manassas, VA, USA). Both cell lines were cultured using Dulbecco’s Modified Eagle Media, supplemented with fetal bovine serum and penicillin/streptomycin at 37 °C, 5% CO2 for production and characterization of rVVs. Sera of P. berghei-infected mice polyclonal antibodies were collected through retro-orbital plexus puncture. Horseradish peroxidase (HRP)-conjugated goat anti-mouse immunoglobulin G (IgG) was purchased from Southern Biotech (Birmingham, AL, USA).
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