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Hrp conjugated goat anti mouse igm or igg

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HRP-conjugated goat anti-mouse IgM or IgG is a secondary antibody used for detection in immunoassays. It is conjugated with horseradish peroxidase (HRP) enzyme, which catalyzes a colorimetric reaction for visualization.

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Lab products found in correlation

Nunc maxisorp, by Thermo Fisher Scientific (2 mentions) Bovine serum albumin (bsa), by Merck Group (1 mentions)

Close spelling variants of this product

HRP-conjugated goat anti-mouse IgM Goat anti-IgM-HRP

4 protocols using hrp conjugated goat anti mouse igm or igg

1

Quantifying CspZ-specific IgM and IgG Titers

Cited 1 time
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Forty-two days post immunization, 100 µL blood was collected from 10 mice via submandibular bleeding to isolate serum. The sera were used to determine the titers of immunoglobulin M or G against CspZ using kinetic ELISA as previously described (30 (link)). In brief, microtiter plate wells were coated with 1 µg of recombinant CspZ. After blocking with 5% BSA (Sigma-Aldrich) in phosphate-buffered saline, 50 µL of mouse serum diluted 1:100, 1:300, 1:900, 1:1,800, 1:3,600, 1:7,200, 1:144,000, or 1: 288,000 was added to each well. HRP-conjugated goat anti-mouse IgM or IgG (1:20,000; Bethyl, Montgomery, TX, USA) and 50 µL of tetramethyl benzidine (TMB) solution (ThermoFisher, Waltham, MA, USA) were subsequently added into the wells, and the binding was detected at 620 nm for 10 cycles of 60 s kinetic intervals with 10 s shaking duration in a Sunrise absorbance ELISA plate reader (Tecan, Männedorf, Switzerland). The greatest maximum slope of optical density/minute per sample was multiplied by the respective serum dilution factor to indicate the antibody titers (arbitrary Unit).
Marcinkiewicz A.L., Lieknina I., Kotelovica S., Yang X., Kraiczy P., Pal U., Lin Y.P, & Tars K. (2018). Eliminating Factor H-Binding Activity of Borrelia burgdorferi CspZ Combined with Virus-Like Particle Conjugation Enhances Its Efficacy as a Lyme Disease Vaccine. Frontiers in Immunology, 9, 181.
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2

ViPS-specific Antibody Quantification

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ViPS-specific IgM and IgG in the blood were measured by coating 96-well microtiter plates (Nunc MaxiSorp; Invitrogen, Carlsbad, CA) with 2 μg/ml ViPS purified from S. Typhi clinical isolate C6524 (47 (link)) in DPBS overnight at room temperature. All plates were washed and blocked with 1% BSA in PBS (pH 7.2) (blocking buffer) for 1 h at room temperature. Blood samples from immunized mice were diluted to 1:25 for IgG detection and 1:50 for IgM detection in blocking buffer, and samples were centrifuged (800 × g for 10 min) and cell-free supernatant was used. The dilutions, 1:25 for IgG and 1:50 for IgM, were chosen after evaluating various serum dilutions within the linear range by ELISA. Bound IgM or IgG was measured using HRP-conjugated goat anti-mouse IgM or IgG (Bethyl Laboratories, Montgomery, TX). Because ViPS-specific mouse IgM and IgG reference standards are not available, and the ViPS-specific Abs in mice are likely to be of oligoclonal nature with varying affinities, the Ag-specific Ab levels in the current study were interpreted as nanogram per microliter “equivalents” using normal mouse serum IgM or IgG standards (Bethyl Laboratories, Montgomery, TX).
Alugupalli A.S., Cravens M.P., Walker J.A., Gulandijany D., Dickinson G.S., Debes G.F., Schifferli D.M., Bäumler A.J, & Alugupalli K.R. (2022). The Lack of Natural IgM Increases Susceptibility and Impairs Anti-Vi Polysaccharide IgG Responses in a Mouse Model of Typhoid. ImmunoHorizons, 6(12), 807-816.
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3

Measuring ViPS-specific IgM and IgG in Mice

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ViPS-specific IgM and IgG were measured by incubating 96-well microtiter plates (Nunc MultiSorp 467340; Nunc A/S, Roskilde, Denmark) with 2 µg/ml of ViPS in DPBS overnight at room temperature. All plates were washed and blocked with 2% Bovine serum albumin (BSA) in PBS pH 7.2 (blocking buffer) for 2 hours at room temperature. Blood from immunized mice was diluted to 1:25 for IgG detection and 1:50 for IgM detection in blocking buffer, samples were centrifuged (800 × g for 10 min.) and cell-free supernatant was used. The dilutions, 1:25 for IgG and 1:50 for IgM were chosen after evaluating various serum dilutions within the linear range by ELISA (data not shown). Bound IgM or IgG was measured using HRP-conjugated goat anti-mouse IgM or IgG (Bethyl Laboratories, Montgomery, TX). Since ViPS-specific mouse IgM and IgG reference standards are not available, and the ViPS-specific antibodies in mice are likely to be of oligoclonal nature with varying affinities, the antigen-specific antibody levels in the present study were interpreted as ng/µl “equivalents” using normal mouse serum IgM or IgG standards (Bethyl Laboratories, Montgomery, TX), as described previously (28 (link)–30 (link)).
Pandya K.D., Palomo-Caturla I., Walker J.A., Sandilya V., Zhong Z, & Alugupalli K.R. (2018). An unmutated IgM response to the Vi Polysaccharide of Salmonella Typhi contributes to protective immunity in a murine model of Typhoid. Journal of immunology (Baltimore, Md. : 1950), 200(12), 4078-4084.
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4

ViPS-specific Antibody Quantification

Check if the same lab product or an alternative is used in the 5 most similar protocols
ViPS-specific IgM and IgG in the blood were measured by coating 96-well microtiter plates (Nunc MaxiSorp; Invitrogen, Carlsbad, CA) with 2 μg/ml ViPS purified from S. Typhi clinical isolate C6524 (47 (link)) in DPBS overnight at room temperature. All plates were washed and blocked with 1% BSA in PBS (pH 7.2) (blocking buffer) for 1 h at room temperature. Blood samples from immunized mice were diluted to 1:25 for IgG detection and 1:50 for IgM detection in blocking buffer, and samples were centrifuged (800 × g for 10 min) and cell-free supernatant was used. The dilutions, 1:25 for IgG and 1:50 for IgM, were chosen after evaluating various serum dilutions within the linear range by ELISA. Bound IgM or IgG was measured using HRP-conjugated goat anti-mouse IgM or IgG (Bethyl Laboratories, Montgomery, TX). Because ViPS-specific mouse IgM and IgG reference standards are not available, and the ViPS-specific Abs in mice are likely to be of oligoclonal nature with varying affinities, the Ag-specific Ab levels in the current study were interpreted as nanogram per microliter “equivalents” using normal mouse serum IgM or IgG standards (Bethyl Laboratories, Montgomery, TX).
Alugupalli A.S., Cravens M.P., Walker J.A., Gulandijany D., Dickinson G.S., Debes G.F., Schifferli D.M., Bäumler A.J, & Alugupalli K.R. (2022). The Lack of Natural IgM Increases Susceptibility and Impairs Anti-Vi Polysaccharide IgG Responses in a Mouse Model of Typhoid. ImmunoHorizons, 6(12), 807-816.
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