Ten days post challenge (p.c.), plasma samples from CB6F1 mice were obtained, and viremia was determined in a focal infectivity assay [56 (link)]. Serial dilutions of plasma were incubated with M. dunni cells for 3 days under standard tissue culture conditions. When cells reached ~100% confluence, they were fixed with ethanol, labeled with F-MuLV Env-specific MAb 720 [48 (link)], and then with a horseradish peroxidase (HRP)-conjugated rabbit antimouse Ig antibody (Dako, Hamburg, Germany). The assay was developed using aminoethylcarbazole (Sigma-Aldrich, Deisenhofen, Germany) as substrate to detect foci. Foci were counted, and focus-forming units (FFU)/ml plasma were calculated.
Horseradish peroxidase hrp conjugated rabbit anti mouse ig antibody
Manufactured by Agilent Technologies
Sourced in Germany
Horseradish peroxidase (HRP)-conjugated rabbit anti-mouse Ig antibody is a laboratory reagent used in various immunoassay techniques. It consists of a polyclonal antibody derived from rabbits that specifically binds to mouse immunoglobulins, conjugated with the enzyme horseradish peroxidase. This conjugate can be used to detect and quantify the presence of mouse antibodies in samples.
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4 protocols using horseradish peroxidase hrp conjugated rabbit anti mouse ig antibody
1
Plasma Viremia Quantification Assay
2
Quantifying Murine Retrovirus Viremia
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Ten days post challenge (p.c.), plasma samples from CB6F1 mice were obtained, and viremia was determined in a focal infectivity assay [56 (link)]. Serial dilutions of plasma were incubated with M. dunni cells for 3 days under standard tissue culture conditions. When cells reached ~100% confluence, they were fixed with ethanol, labeled with F-MuLV Env-specific MAb 720 [49 (link)], and then with a horseradish peroxidase (HRP)-conjugated rabbit anti-mouse Ig antibody (Dako, Hamburg, Germany). The assay was developed using aminoethylcarbazole (Sigma-Aldrich, Deisenhofen, Germany) as substrate to detect foci. Foci were counted, and focus-forming units (FFU)/ml plasma were calculated.
3
Viral Load Quantification by IC Assay
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Determination of viral loads by IC assay was performed as described previously (36 (link)). Spleens were cropped and rinsed with RPMI containing 10% FCS and 50 µg mL−1 penicillin/streptomycin. Spleen cells were counted and serially diluted before seeding them onto Mus dunni tail fibroblast cells and incubated under standard tissue culture conditions for 3 days, fixed with ethanol and labeled with the primary F-MuLV Env-specific MAb 720 (37 (link)). After washing, a secondary horseradish peroxidase (HRP)-conjugated rabbit anti-mouse Ig antibody (Dako) was added. Foci representing ICs were detectable after adding of aminoethylcarbazole (Sigma-Aldrich) as substrate for HRP. Foci were counted and ICs/spleen values were calculated.
4
Quantifying Retroviral Infectious Centers
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Determination of viral loads by infectious-center assay was performed as described previously (55 (link)). Spleens were cropped and rinsed with RPMI medium containing 10% FCS and 50 μg ml−1 penicillin/streptomycin. Spleen cells were counted and serially diluted before they were seeded onto Mus dunni cells, and then they were incubated under standard tissue culture conditions for 3 days, fixed with ethanol, and labeled with the primary F-MuLV Env-specific monoclonal antibody 720 (56 (link)). After the cells were washed, a secondary horseradish peroxidase (HRP)-conjugated rabbit anti-mouse Ig antibody (Dako) was added. Foci representing infectious centers were detectable after the addition of aminoethylcarbazole (Sigma-Aldrich) as the substrate for HRP. Foci were counted and numbers of infectious centers (IC) per spleen were calculated.
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