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Anti mouse iga hrp

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Anti-mouse IgA-HRP is a laboratory reagent used for the detection and quantification of mouse immunoglobulin A (IgA) in various applications. It consists of an anti-mouse IgA antibody conjugated to the enzyme horseradish peroxidase (HRP).

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Lab products found in correlation

Anti mouse igg hrp, by Southern Biotech (5 mentions) Anti mouse igg1 hrp, by Southern Biotech (5 mentions) Anti mouse igg2c hrp, by Southern Biotech (4 mentions) 1 step ultra tmb elisa, by Thermo Fisher Scientific (1 mentions) Microplate reader, by Bio-Rad (1 mentions) Anti mouse igg2a hrp, by Southern Biotech (1 mentions) Anti human iga hrp, by Southern Biotech (1 mentions) Anti human igg hrp, by Southern Biotech (1 mentions) Anti mouse ige, by BD (1 mentions) Biotin anti mouse ige, by BD (1 mentions) Tmb elisa substrate solution, by Thermo Fisher Scientific (1 mentions) Streptavidin hrp, by Rockland Immunochemicals (1 mentions) Anti mouse igm hrp, by Southern Biotech (1 mentions) High binding eia ria 96 well plate, by Corning (1 mentions) Vac nacga23, by InvivoGen (1 mentions) Vac pova, by InvivoGen (1 mentions) Mouse ifn γ elisa kit, by Thermo Fisher Scientific (1 mentions) Elisa kit, by Thermo Fisher Scientific (1 mentions)

9 protocols using anti mouse iga hrp

1

SARS-CoV-2 Spike Protein ELISA

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Purified antigens (S or RBD) were coated onto 96-well Maxisorp clear plates at 2 μg/mL in 50 mM Na2CO3 pH 9.6 (70 μL) overnight at 4°C. Coating buffers were aspirated, and wells were blocked with 200 μL of 1X PBS + 0.05% Tween-20 + 1% BSA + 0.02% NaN3 (Blocking buffer, PBSTBA) overnight at 4°C. Heat-inactivated serum samples were diluted in PBSTBA in a separate 96-well polypropylene plate. The plates then were washed thrice with 1X PBS + 0.05% Tween-20 (PBST), followed by addition of 50 μL of respective serum dilutions. Sera were incubated in the blocked ELISA plates for at least 1 h at room temperature. The ELISA plates were again washed thrice in PBST, followed by addition of 50 μL of 1:1,000 anti-mouse IgG-HRP (Southern Biotech Cat. #1030-05) in PBST or 1:1000 of anti-mouse IgA-HRP in PBSTBA (SouthernBiotech). Plates were incubated at room temperature for 1 h, washed thrice in PBST, and then 100 μL of 1-Step Ultra TMB-ELISA was added (ThermoFisher Cat. #34028). Following a 10 to 12-min incubation, reactions were stopped with 50 μL of 2 M sulfuric acid. Optical density (450 nm) measurements were determined using a microplate reader (Bio-Rad).
Hassan A.O., Shrihari S., Gorman M.J., Ying B., Yaun D., Raju S., Chen R.E., Dmitriev I.P., Kashentseva E., Adams L.J., Mann C., Davis-Gardner M.E., Suthar M.S., Shi P.Y., Saphire E.O., Fremont D.H., Curiel D.T., Alter G, & Diamond M.S. (2021). An intranasal vaccine durably protects against SARS-CoV-2 variants in mice. Cell Reports, 36(4), 109452.
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2

SARS-CoV-2 Antibody Quantification in Mice and Humans

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A semi-quantitative ELISA was used to determine the levels of IgG, IgG1, IgG2a, and IgA antibodies in the sera of vaccinated mice. Also, the levels of IgG and IgA antibodies were determined in the sera of recovered COVID-19 patients.
First, high binding ELISA plates (Biomat, Italy) were coated with SARS-CoV-2 S- protein recombinant antigen (Sigma-Aldrich) at 1 μgmL−1. After washing plates, 50 μL of diluted serum samples collected from immunized mice and recovered COVID-19 patients were separately added to wells. After incubation for 1 h at 37 °C, plates were washed with PBS and then 100 μL of the following secondary antibody was separately added: 1) anti-mouse IgG-HRP, 2) anti-mouse IgG1-HRP, 3) anti-mouse IgG2a-HRP, 4) anti-mouse IgA-HRP, 5) anti-human IgG-HRP, 6) anti-human IgA-HRP (Southern Biotech). After incubation and washing with PBS, 50 μL of 3,3′, 5,5′-tetramethylbenzidine was added and then the reactions was stopped by adding 50 μL of 10% sulfuric acid. Finally, the absorbance of each well was read by a Spectrophotometer at 450 nm (BioTek Industries) and serum levels of antibodies was measured using a standard curve.
Mohammadi G., Sotoudehnia Koranni Z, & Jebali A. (2021). The oral vaccine based on self-replicating RNA lipid nanoparticles can simultaneously neutralize both SARS-CoV-2 variants alpha and delta. International Immunopharmacology, 101, 108231.
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3

Enzyme Immunoassay for Antibody Detection

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High-binding EIA/RIA 96-well plates (Costar) were coated overnight with 10µg/mL BSA-NP32 or BSA-NP5 (Biosearch Technologies Inc) diluted in ELISA coating buffer (28.6mM Na2Co3, 11.9mM NaHCO3, pH 9.6), or 2µg/mL anti-mouse IgE (BD 553413) diluted in PBS. Wells were subsequently washed four times with 0.05% Tween20/PBS, then blocked with 3% BSA/PBS for 2 hours at room temperature. After washing, sera was serially diluted in 1% BSA/PBS, added to wells and incubated for 2 hours at room temperature. Wells were washed and incubated with anti-mouse IgG-HRP (1030-05), anti-mouse IgM-HRP (1021-05), anti-mouse IgA-HRP (1040-05) from Southern Biotech, or anti-mouse IgE-biotin (BD 553419) diluted in 1% BSA/PBS for 2 hours at room temperature. Biotinylated antibodies were further incubated with streptavidin-HRP (Rockland) for 40 minutes at room temperature. After washing, HRP was detected with 1X TMB ELISA Substrate Solution (eBioscience) and color development was stopped with 1M orthophosphoric acid. Plates were analyzed at 450nm with a Biotrak II plate reader.
Bastow C.R., Kara E.E., Tyllis T.S., Vinuesa C.G., McColl S.R, & Comerford I. (2022). TFR Cells Express Functional CCR6 But It Is Dispensable for Their Development and Localization During Splenic Humoral Immune Responses. Frontiers in Immunology, 13, 873586.
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4

Quantification of Anti-OVA Antibodies

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The Ag-specific Abs were determined by ELISA. The anti-IgG-HRP used was anti-mouse IgG1-HRP (cat#1070-05; Southern Biotech, Birmingham, AL), anti-mouse IgG2C-HRP (cat#1079-05; Southern Biotech), and anti-mouse IgA-HRP (cat#1040-05; Southern Biotech). Total anti-OVA IgG2A, IgG1 and IgA were quantified using a mouse anti-OVA IgG2A kit (cat#3015; Chondrex, Redmond, WA), anti-OVA IgG1 kit (cat#3013; Chondrex, Redmond, WA), and anti-OVA IgA kit (cat#3018; Chondrex, Redmond, WA).
Blaauboer S.M., Mansouri S., Tucker H.R., Wang H.L., Gabrielle V.D, & Jin L. (2015). The mucosal adjuvant cyclic di-GMP enhances antigen uptake and selectively activates pinocytosis-efficient cells in vivo. eLife, 4, e06670.
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5

Intranasal Vaccination with CDG Adjuvant

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Groups of mice were intranasally vaccinated with CDG (5μg) adjuvanted antigen (2μg) or antigen alone (26 (link)). For intranasal vaccination, animals were anesthetized using isoflurane in an E-Z Anesthesia system (Euthanex Corp, Palmer, PA). Antigen, with or without CDG was administered in 30μl saline. Sera were collected at the indicated time points after the last immunization. The antigen-specific Abs were determined by ELISA. Secondary Abs used were anti-mouse IgG-HRP (Southern Biotech, 1033– 05), and anti-mouse IgA-HRP (Southern Biotech, 1040–05).To determine Ag-specific TH response, splenocytes and lung cells from antigen or CDG + antigen immunized mice were stimulated with 5μg/ml antigen for four days in culture. TH cytokines were measured in the supernatant by ELISA.
Mansouri S., Katikaneni D.S., Gogoi H, & Jin L. (2021). moDCs differentiate into Bcl6+ mature moDCs to promote cyclic di-GMP vaccine adjuvant-induced memory TH cells in the lung. Journal of immunology (Baltimore, Md. : 1950), 206(9), 2233-2245.
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6

Intranasal Vaccination Enhances Mucosal Immunity

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Groups of mice (4 per group) were intranasally vaccinated with 5μg 2′3′-cGAMP adjuvanted PspA (2μg, BEI Resources) or PspA alone(11 ). Mice were immunized twice at 14 days interval. For intranasal vaccination, animals were anesthetized using isoflurane in an E–Z Anesthesia system (Euthanex Corp, Palmer, PA). PspA, with or without 2′3-cGAMP was administered in 20μl saline. Sera, BALF, and nasal washes were collected 14 days after the last immunization. The PspA-specific Abs were determined by ELISA. Secondary Abs used were anti-mouse IgG1-HRP (Southern Biotech, cat#1070-05), anti-mouse IgG2C-HRP (Southern Biotech, cat#1079-05), and anti-mouse IgA-HRP (Southern Biotech, cat#1040-05). To determine Ag-specific Th response, splenocytes from PspA or 2′3-cGAMP + PspA immunized mice were stimulated with 5μg/ml PspA for four days in culture. Th1, Th2, and Th17 cytokines were measured in the supernatant by ELISA.
Patel S., Blaauboer S.M., Tucker H.R., Mansouri S., Ruiz-Moreno J.S., Hamann L., Schumann R.R., Opitz B, & Jin L. (2016). The common R71H-G230A-R293Q (HAQ) human TMEM173 is a null allele. Journal of immunology (Baltimore, Md. : 1950), 198(2), 776-787.
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7

Intranasal Vaccination: PspA with CDG

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Groups of mice (4 per group) were intranasally vaccinated with CDG (5µg, Invivogen, cat# vac-cdg) adjuvanted PspA (2µg, BEI Resources) or PspA alone 9 . Mice were immunized twice at 14 days interval. For intranasal vaccination, animals were anesthetized using isoflurane in an E-Z Anesthesia system (Euthanex Corp, Palmer, PA). PspA, with or without CDG was administered in 20µl saline. Secondary Abs used were anti-mouse IgG1-HRP (Southern Biotech, cat#1070– 05), anti-mouse IgG2C-HRP (Southern Biotech, cat#1079–05), and anti-mouse IgA-HRP (Southern Biotech, cat#1040–05).
Mansouri S., Patel S., Katikaneni D.S., Blaauboer S.M., Wang W., Schattgen S., Fitzgerald K, & Jin L. (2018). Immature lung TNFR2− conventional DC 2 subpopulation activates moDCs to promote cyclic di-GMP mucosal adjuvant responses in vivo. Mucosal immunology, 12(1), 277-289.
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8

Intranasal 2'3'-cGAMP Adjuvanted OVA Immunization

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Groups of mice (4 mice per group) were immunized i.n. with 5μg 2’3′-cGAMP (Invivogen, vac-nacga23) adjuvanted OVA (Invivogen, vac-pova) (2μg) or OVA alone. Mice were immunized twice at a 14 days interval. Sera, BALF was collected 28 days after the last immunization. The OVA-specific Abs were determined by ELISA. Secondary Abs used were anti-mouse IgG-HRP or anti-mouse IgA-HRP (SouthernBiotech, 1040–05, 7040). To determine the Ag-specific memory TH response, splenocytes from OVA or 2′3-cGAMP + OVA immunized mice were stimulated with 5 μg/ml OVA for 4 days in culture. Th1, Th2, and Th17 cytokines were measured in the supernatant by ELISA kit from ThermoFisher (mouse IFNγ ELISA kit, 88-7314-22; mouse IL-17a ELISA kit, 88-8711-22; mouse IL-13 ELISA kit, 88-7137-22).
Mansouri S., Gogoi H., Patel S., Katikaneni D.S., Singh A., Aybar-Torres A., de Lartigue G, & Jin L. (2022). MPYS Modulates Fatty Acid Metabolism and Immune Tolerance at Homeostasis Independent of Type I IFNs. The Journal of Immunology Author Choice, 209(11), 2114-2132.
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9

Intranasal Vaccination: PspA with CDG

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Groups of mice (4 per group) were intranasally vaccinated with CDG (5µg, Invivogen, cat# vac-cdg) adjuvanted PspA (2µg, BEI Resources) or PspA alone 9 . Mice were immunized twice at 14 days interval. For intranasal vaccination, animals were anesthetized using isoflurane in an E-Z Anesthesia system (Euthanex Corp, Palmer, PA). PspA, with or without CDG was administered in 20µl saline. Secondary Abs used were anti-mouse IgG1-HRP (Southern Biotech, cat#1070– 05), anti-mouse IgG2C-HRP (Southern Biotech, cat#1079–05), and anti-mouse IgA-HRP (Southern Biotech, cat#1040–05).
Mansouri S., Patel S., Katikaneni D.S., Blaauboer S.M., Wang W., Schattgen S., Fitzgerald K, & Jin L. (2018). Immature lung TNFR2− conventional DC 2 subpopulation activates moDCs to promote cyclic di-GMP mucosal adjuvant responses in vivo. Mucosal immunology, 12(1), 277-289.
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