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Hrp conjugated goat anti guinea pig secondary antibody

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The HRP-conjugated goat anti-guinea pig secondary antibody is a laboratory reagent used to detect and visualize the presence of guinea pig primary antibodies in various immunoassays. This secondary antibody is conjugated with horseradish peroxidase (HRP), an enzyme that catalyzes a colorimetric or chemiluminescent reaction, allowing for the detection and quantification of the target antibodies.

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Lab products found in correlation

Spectramax plus elisa plate reader, by Molecular Devices (3 mentions) Elisa plate, by Thermo Fisher Scientific (2 mentions) Hrp conjugated goat anti human igg secondary antibody, by Thermo Fisher Scientific (1 mentions) Maxisorp elisa plate, by Thermo Fisher Scientific (1 mentions) Tween 20, by Merck Group (1 mentions)

Close spelling variants of this product

HRP-conjugated goat anti-guinea pig HRP-conjugated anti-guinea pig antibody Goat anti-guinea pig-HRP HRP-conjugated secondary antibodies Secondary HRP antibodies An anti-HRP antibody

3 protocols using hrp conjugated goat anti guinea pig secondary antibody

1

Serum Antibodies against HIV Antigens

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Serum binding antibodies against gp140 foldon and V1/V2 scaffolds were measured by endpoint enzyme-linked immunosorbant assays (ELISAs) as described previously (Nkolola et al., 2010 (link)). Briefly, ELISA plates (Thermo Scientific) were coated with individual gp140s or V1/V2 scaffolds (Immune Technology) and incubated overnight. Guinea pig sera were then added in serial dilutions and later detected with an HRP-conjugated goat anti-guinea pig secondary antibody (Jackson ImmunoResearch Laboratories). Plates were developed and read using the Spectramax Plus ELISA plate reader (Molecular Devices) and Softmax Pro-4.7.1 software. End-point titers were considered positive at the highest dilution that maintained an absorbance >2-fold above background values.
Bricault C.A., Yusim K., Seaman M.S., Yoon H., Theiler J., Giorgi E.E., Wagh K., Theiler M., Hraber P., Macke J.P., Kreider E.F., Learn G.H., Hahn B.H., Scheid J.F., Kovacs J.M., Shields J.L., Lavine C.L., Ghantous F., Rist M., Bayne M.G., Neubauer G.H., McMahan K., Peng H., Chéneau C., Jones J.J., Zeng J., Ochsenbauer C., Nkolola J.P., Stephenson K.E., Chen B., Gnanakaran S., Bonsignori M., Williams L.D., Haynes B.F., Doria-Rose N., Mascola J.R., Montefiori D.C., Barouch D.H, & Korber B. (2019). HIV-1 Neutralizing Antibody Signatures and Application to Epitope-Targeted Vaccine Design. Cell Host & Microbe, 25(1), 59-72.e8.
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2

Serum Antibody Avidity Measurement

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Serum binding antibody avidities to HIV-1 Env gp140 were measured by a urea disruption enzyme-linked immunosorbent assay (ELISA) (58 (link)). Briefly, ELISA plates (Thermo Scientific) were coated with individual Env gp140s and incubated overnight. Guinea pig sera were prepared in a dilution plate to values between 1.0 and 1.5 at optical densities of 450 nm (OD450) to OD800 to provide antibody concentrations within a linear range. Sera were plated in duplicate twice; one duplicate was treated with 8 molar urea (Sigma-Aldrich) and the other with distilled water as a control. Plates were then incubated with an HRP-conjugated goat anti-guinea pig secondary antibody (Jackson ImmunoResearch Laboratories). Plates were developed and read using the SpectraMax plus ELISA plate reader (Molecular Devices) and SoftMax Pro 4.7.1 software (OD450 to OD800). Percent avidity was calculated using the following formula: [(average absorbance of urea-treated sample/average absorbance of water-treated, matched sample) × 100]. The avidity index describes 0 to 30% as low-avidity, 30 to 50% as moderate-avidity, and >50% as high-avidity binding antibodies.
Bricault C.A., Kovacs J.M., Badamchi-Zadeh A., McKee K., Shields J.L., Gunn B.M., Neubauer G.H., Ghantous F., Jennings J., Gillis L., Perry J., Nkolola J.P., Alter G., Chen B., Stephenson K.E., Doria-Rose N., Mascola J.R., Seaman M.S, & Barouch D.H. (2018). Neutralizing Antibody Responses following Long-Term Vaccination with HIV-1 Env gp140 in Guinea Pigs. Journal of Virology, 92(13), e00369-18.
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3

Serum Antibody Titer Assay for Clade C gp140

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Serum binding antibody titers against clade C gp140 trimer were determined by endpoint ELISAs as previously described [11 (link), 12 (link)]. Briefly, 96-well Maxisorp ELISA plates (Thermo Fisher Scientific), coated over-night with 100 μl/well of 1 μg/ml clade C gp140, were blocked for 2.5 hours with PBS/Casein (Pierce). Guinea pig sera were then added in serial dilutions and incubated for 1 hour at room temperature. The plates were washed three times with PBS containing 0.05% Tween 20 (Sigma) and incubated for 1 hour with a 1/2,000 dilution of a horseradish peroxidase (HRP)-conjugated goat anti-guinea pig secondary antibody (Jackson ImmunoResearch Laboratories) or HRP-conjugated goat anti-human IgG secondary antibody (Fisher Scientific) for NHP assays. The plates were washed three times and developed with SureBlue tetramethylbenzidine (TMB) microwell peroxidase (KPL Research Products), stopped by the addition of stop solution (KPL Research products), and analyzed at 450 nm/550 nm with a Spectramax Plus ELISA plate reader (Molecular Devices) using Softmax Pro 4.7.1 software. ELISA endpoint titers were defined as the highest reciprocal serum dilution that yielded an absorbance >2-fold over background values.
Nkolola J.P., Cheung A., Perry J.R., Carter D., Reed S., Schuitemaker H., Pau M.G., Seaman M.S., Chen B, & Barouch D.H. (2014). Comparison of multiple adjuvants on the stability and immunogenicity of a clade C HIV-1 gp140 trimer. Vaccine, 32(18), 2109-2116.
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