Ninety six-well EIA/RIA plate wells were filled with 100 µl of heat-inactivated purified HSV-2 (104 to 105 pfu equivalent per 100 µl) or heat-inactivated purified VSV (5 × 105 pfu equivalent per 100 µl) for virus-specific Ig measurement or goat anti-mouse Ig (1:1000; SouthernBiotech, 1010-01) for total Ig measurement in carbonate buffer (pH 9.5) and then incubated overnight at 4°C. On the following day, these plates were washed with PBS-Tween 20 and blocked 2hr with 5% FBS in PBS. Tissue samples and serum samples in ABC buffer were then plated in the wells and incubate for at least four hours at ambient temperature. After washing in PBS-Tween 20, HRP-conjugated anti-mouse IgG1, IgG3, IgM, IgA, IgG2a, IgG2b or IgG2c (SouthernBiotech) was added in the wells for 1 h, followed by washing and adding TMB solution (eBioscience). Reactions were stopped with 1N H2SO4 and absorbance was measured at 450 nm. The sample Ab titers were defined by using Ig standard (C57BL/6 Mouse Immunoglobulin Panel; SouthernBiotech) or mouse IgG2a (HOPC-1; SouthernBiotech).
Hopc 1
Manufactured by Southern Biotech
The HOPC-1 is a high-output protein concentrator designed for efficient separation and purification of proteins from complex biological samples. It utilizes a high-performance tangential flow filtration system to concentrate protein solutions while maintaining the integrity of the target molecules.
Automatically generated - may contain errors
2 protocols using hopc 1
1
ELISA for Virus-specific Antibodies
2
ELISA for Virus-specific Antibodies
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Ninety six-well EIA/RIA plate wells were filled with 100 µl of heat-inactivated purified HSV-2 (104 to 105 pfu equivalent per 100 µl) or heat-inactivated purified VSV (5 × 105 pfu equivalent per 100 µl) for virus-specific Ig measurement or goat anti-mouse Ig (1:1000; SouthernBiotech, 1010-01) for total Ig measurement in carbonate buffer (pH 9.5) and then incubated overnight at 4°C. On the following day, these plates were washed with PBS-Tween 20 and blocked 2hr with 5% FBS in PBS. Tissue samples and serum samples in ABC buffer were then plated in the wells and incubate for at least four hours at ambient temperature. After washing in PBS-Tween 20, HRP-conjugated anti-mouse IgG1, IgG3, IgM, IgA, IgG2a, IgG2b or IgG2c (SouthernBiotech) was added in the wells for 1 h, followed by washing and adding TMB solution (eBioscience). Reactions were stopped with 1N H2SO4 and absorbance was measured at 450 nm. The sample Ab titers were defined by using Ig standard (C57BL/6 Mouse Immunoglobulin Panel; SouthernBiotech) or mouse IgG2a (HOPC-1; SouthernBiotech).
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