Plates were pre-coated with Goat anti-Mouse Ig (Southern Biotech), blocked with PBS-BB (PBS + 0.05% Tween 20 + 1% BSA) and serum was applied. HRP conjugated secondary antibodies anti-IgA-HRP, anti-IgG3-HRP, anti-IgG1-HRP, anti-IgM-HRP, anti-IgG2b-HRP, and anti-IgG-HRP (all from Southern Biotech) were applied and subsequently exposed with a slow kinetic TMB solution (Sigma). The reaction was terminated with 1N HCL and absorbance was measured at 450 nm on a SpectraMax 340 PC plate reader and analyzed using SoftMax Pro 4.8 Software.
Anti igg3 hrp
Manufactured by Southern Biotech
Anti-IgG3-HRP is a laboratory reagent used for the detection and quantification of immunoglobulin G subclass 3 (IgG3) in various biological samples. It consists of an antibody specific to the IgG3 molecule conjugated with the enzyme horseradish peroxidase (HRP). This conjugate can be utilized in immunoassay techniques, such as ELISA, to measure IgG3 levels.
Automatically generated - may contain errors
Lab products found in correlation
Anti igm hrp, by Southern Biotech (4 mentions)
Anti igg1 hrp, by Southern Biotech (4 mentions)
Anti igg2b hrp, by Southern Biotech (4 mentions)
Slow kinetic tmb solution, by Merck Group (2 mentions)
Goat anti mouse ig, by Southern Biotech (2 mentions)
Recombinant mouse baff, by R&D Systems (1 mentions)
Tmb substrate, by BioLegend (1 mentions)
Igg2b, by Merck Group (1 mentions)
Anti iga biotin, by Southern Biotech (1 mentions)
Streptavidin hrp, by Southern Biotech (1 mentions)
Igg2c, by Southern Biotech (1 mentions)
Igg2b, by Southern Biotech (1 mentions)
4 protocols using anti igg3 hrp
1
Isotype-Specific ELISA Protocol
2
Isotype-Specific ELISA Protocol
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Plates were pre-coated with Goat anti-Mouse Ig (Southern Biotech), blocked with PBS-BB (PBS + 0.05% Tween 20 + 1% BSA) and serum was applied. HRP conjugated secondary antibodies anti-IgA-HRP, anti-IgG3-HRP, anti-IgG1-HRP, anti-IgM-HRP, anti-IgG2b-HRP, and anti-IgG-HRP (all from Southern Biotech) were applied and subsequently exposed with a slow kinetic TMB solution (Sigma). The reaction was terminated with 1N HCL and absorbance was measured at 450 nm on a SpectraMax 340 PC plate reader and analyzed using SoftMax Pro 4.8 Software.
3
Antibody and BAFF Quantification by ELISA
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ELISA was used to detect antigen-specific IgM, IgG1, IgG2b, and IgG3 titers by capturing serum antibodies using NIP11-OVA (Jackson ImmunoResearch, Inc) coated plates (NP-specific) or methylated BSA and calf thymus DNA (Sigma-Aldrich) coated plates (DNA-specific), as described previously (30 (link), 31 (link)). Bound antibody was detected with anti-IgM-HRP, anti-IgG1-HRP, anti-IgG2b-HRP or anti-IgG3-HRP (Southern Biotech), TMB substrate (Biolegend), and read on a Molecular Devices microplate reader at 450nm. Titer was determined to be lowest serum dilution achieving signal greater than 2x background. BAFF in sera was measured by a sandwich ELISA using BAFF- specific clones 5A8 (capture) and 1C9-biotin (detection) (Axxora) as performed previously (31 (link)). Recombinant mouse BAFF (R&D Systems) served as standard.
4
Serum Antibody Isotype Analysis in Mice
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Blood for serum antibody analysis was collected from all mice at 49 days of age. Plates were coated with anti-mouse IgM, IgG1, IgG2b, IgG2c, IgG3 or IgA (Southern Biotech, Birmingham, AL, USA) for 24 h before the addition of diluted serum. IgM, IgG1, IgG2b, IgG3 standards were obtained from Sigma-Aldrich, IgG2c standard was from Southern Biotech, and IgA standard was from Organon Tekcika–Cappel (Durham, NC, USA). Serum Ig was detected with anti-IgM-HRP, anti-IgG1-HRP, anti-IgG2b-HRP, anti-IgG2c-HRP, anti-IgG3-HRP, anti-IgA-biotin and streptavidin-HRP (Southern Biotech) and visualised using ABTS substrate (2,2′-Azinobis (3-ethylbenzthiazoline Sulfonic Acid); Sigma-Aldrich). All samples and standards were measured in duplicate.
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