Analyses of induced antibodies in mice were performed mainly as previously described by Vallejo et al32 (link). p210, p210-PADRE, or PADRE peptides were used for coating (20 μg/mL of each in PBS) microtiter plates (Nunc MaxiSorp, Nunc) over night at 4 °C. Coated plates were washed with PBS with 0.05% Tween-20 and thereafter blocked with SuperBlock™ in Tris-buffered saline (Pierce) during 30 min at room temperature followed by an incubation of mouse plasma diluted 1:1000 in PBS-0.05% Tween-20 for 2 h at RT. After rinsing, bound antibodies were detected using alkaline phosphatase–conjugated goat anti-mouse IgG1 (BD Biosciences), IgG2c antibodies (Southern Biotech), or IgM antibodies (Vector Laboratories), which were incubated for 2 h at room temperature, and a color reaction was developed using phosphatase substrate kit (Pierce). The absorbance (405 nm) was measured and background absorbance was subtracted. Specificity assay of apoB100 p210-induced antibodies were performed by a competition ELISA. p210 peptide were coated in microtiter wells and binding of antibodies in plasma from p210-PADRE- or PADRE-immunized mice (dilution 1:1000) were competed by increasing concentrations of p210 peptide in solution (0, 1, 10 or 100 µg/ml). Bound antibodies were detected as described above.
Igg2c antibodies
Manufactured by Southern Biotech
IgG2c antibodies are a type of immunoglobulin G (IgG) antibody found in mammals. They play a role in the immune system's response to various antigens. IgG2c antibodies are involved in the neutralization and clearance of pathogens, as well as the activation of the complement system.
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Lab products found in correlation
2 protocols using igg2c antibodies
1
Induced Antibody Analysis in Mice
2
ELISA for Malaria Antibody Detection
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ELISA for Detection of Malaria-Specific Antibodies Microtiter plates were coated with recombinant P. berghei ANKA MSP-1 19 (1 mg/ml) in carbonate buffer pH 9.6 by overnight incubation at 4 C. Empty sites were blocked with 5% skim milk for 1 h at 37 C. After washing with 0.05% Tween 20 in PBS, plates were incubated with antisera in serial dilutions for 1 h at 37 C. The plates were washed and incubated with a peroxidase-conjugated rabbit anti-mouse antibody (Pierce, Rockford, IL, USA). Isotype titers were determined after incubation with anti-IgG 1 (Pierce, ThermoFisher Scientific) or -IgG 2c antibodies (Southern Biotech), followed by incubation with a peroxidase-conjugated goat anti-rabbit antibody (Pierce, Rockford, IL). Bound complexes were detected by reaction with tetramethy-benzidine (KBL, Maryland) and H 2 O 2 . Absorbance was read at 450 nm on a Chameleon plate reader (Hidex). To determine the relative affinity of antibodies, plates were coated with different concentration of recombinant P. berghei ANKA MSP-1 19 (0.1-40 mg/ml) in carbonate buffer pH 9.6. After blocking, immune sera were added at a 1:50 dilution followed by incubation with peroxidase-conjugated rabbit anti-mouse antibody, specific for all IgG subclasses (Sigma). Bound complexes were detected as described above. The relative avidity was estimated by applying non-linear models to calculate EC 50 values.
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