Hrp conjugated goat anti mouse igm

Serum levels of anti-PEG IgG and IgM were detected on day 7 after the first and second dose of mRNA vaccine, while anti-PEG IgE was detected 24 h after vaccination. 96-well plates were pre-coated with 1 μg PEGfilgrastim and sealed with 1 % BSA solution for 1 h. After washing the plates three times, diluted mouse serum was added to the wells and incubated at 37 °C for 1.5 h. Mouse serum was diluted by factors of 100, 20, and 10 times for measuring anti-PEG IgM, anti-PEG IgG, and anti-PEG IgE, respectively. After washing the plate 5 times, HRP-conjugated goat anti-mouse IgM (Invitrogen™) at a dilution of 1/2000, HRP-conjugated anti-mouse IgG (Invitrogen™) at a dilution of 1/5000, and HRP-conjugated anti-mouse IgE (Invitrogen™) at a dilution of 1/1000 were added to the plates and incubated at 37 °C for 1 h each. Following another round of washing five times, color development was achieved using TMB for 15 min before being terminated with a solution containing sulfuric acid at a concentration of 1 mol/L. OD values were measured at wavelengths of both450 nm and630 nm using a multifunctional plate reader.

Herculase II Fusion DNA
polymerase was obtained from Agilent (Seoul, Korea). Oligonucleotides
were obtained from Bionics (Seoul, Korea). The plasmid miniprep kit
was obtained from GeneAll Biotechnology (Seoul, Korea). Protein A
beads were obtained from GE healthcare (Piscataway, NJ). A disposable
gravity column was obtained from Bio-Rad (Daejeon, Korea). Ultrafiltration
devices were obtained from Millipore (centrifugal filter tube Ultra-4,
MWCO 3k; Seoul, Korea). The 96-well Maxi-binding ELISA plate was obtained
from SPL (Seoul, Korea). Freestyle 293 medium was obtained from Gibco
(Waltham, MA). PEI was obtained from Polysciences (Warrington, PA).
LTA was obtained from Sigma (Seoul, Korea). Goat anti-mouse IgG HRP
was obtained from Pierce (Seoul, Korea). Mouse anti-S. aureus monoclonal IgM and rabbit anti-S. aureus polyclonal IgG were obtained from Thermo.
HRP-conjugated goat anti-rabbit IgG (H+L) and HRP-conjugated goat
anti-mouse IgM were obtained from Invitrogen (Waltham, MA). Other
chemicals and reagents, unless otherwise indicated, were from Sigma.