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Anti human igm igg iga

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Anti-human IgM+IgG+IgA is a laboratory reagent that can detect and measure the presence of human immunoglobulins IgM, IgG, and IgA in various samples. It is commonly used in immunological assays and tests.

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Lab products found in correlation

Rpa t4, by Thermo Fisher Scientific (2 mentions) Facscanto, by BD (2 mentions) Alexa fluor 647, by Jackson ImmunoResearch (2 mentions) Horseradish peroxidase hrp conjugated abs able to detect the fc region of human igg, by Thermo Fisher Scientific (2 mentions) Interleukin 4 (il 4), by Thermo Fisher Scientific (1 mentions) Dynabeads untouched human b cells kit, by Thermo Fisher Scientific (1 mentions) Scd40l, by Enzo Life Sciences (1 mentions) Anti cd3 ucht1, by Thermo Fisher Scientific (1 mentions) Cd8 sk1, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

Anti-human IgM (9 citations) Anti-human IgA (3 citations) Human IgM (2 citations) Peroxidase-conjugated fragment goat anti-human IgA or IgG or IgM antibodies (2 citations) F(ab’)2 fragment goat anti-human IgA+IgG+IgM (H+L) (2 citations) HRP-conjugated goat anti-human IgA/IgG/IgM (2 citations) F(ab)2 anti-human IgM/IgG/IgA (2 citations) AffiniPure Goat Anti-Human IgA+IgG+IgM (H+L), (2 citations) An affiniPure F (ab’) Fragment Goat Anti-Human IgA/ IgG/IgM (H + L) (2 citations)

11 protocols using anti human igm igg iga

1

SARS-CoV-2 Antibody Detection Assay Protocol

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Plasma from SARS-CoV-2 naive and PI donors were collected, heat-inactivated for 1 h at 56°C and stored at −80°C until ready to use in subsequent experiments. Plasma from uninfected donors collected before the pandemic were used as negative controls and used to calculate the seropositivity threshold in our ELISA, cell-based ELISA, ADCC and flow cytometry assays (see below). The RBD-specific monoclonal antibody CR3022 was used as a positive control in our ELISA, cell-based ELISA, and flow cytometry assays and was previously described (Anand et al., 2021a (link); Beaudoin-Bussières et al., 2020 ; ter Meulen et al., 2006 (link); Prévost et al., 2020 (link)). Horseradish peroxidase (HRP)-conjugated Abs able to detect all Ig isotypes (anti-human IgM+IgG+IgA; Jackson ImmunoResearch Laboratories) or specific for the Fc region of human IgG (Invitrogen), the Fc region of human IgM (Jackson ImmunoResearch Laboratories) or the Fc region of human IgA (Jackson ImmunoResearch Laboratories) were used as secondary Abs to detect Ab binding in ELISA and cell-based ELISA experiments. Alexa Fluor-647-conjugated goat anti-human Abs able to detect all Ig isotypes (anti-human IgM+IgG+IgA; Jackson ImmunoResearch Laboratories) were used as secondary Ab to detect plasma binding in flow cytometry experiments.
Tauzin A., Gong S.Y., Beaudoin-Bussières G., Vézina D., Gasser R., Nault L., Marchitto L., Benlarbi M., Chatterjee D., Nayrac M., Laumaea A., Prévost J., Boutin M., Sannier G., Nicolas A., Bourassa C., Gendron-Lepage G., Medjahed H., Goyette G., Bo Y., Perreault J., Gokool L., Morrisseau C., Arlotto P., Bazin R., Dubé M., De Serres G., Brousseau N., Richard J., Rovito R., Côté M., Tremblay C., Marchetti G.C., Duerr R., Martel-Laferrière V., Kaufmann D.E, & Finzi A. (2021). Strong humoral immune responses against SARS-CoV-2 Spike after BNT162b2 mRNA vaccination with a 16-week interval between doses. Cell Host & Microbe, 30(1), 97-109.e5.
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2

SARS-CoV-2 Plasma Antibody Detection

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Plasma from SARS-CoV-2-infected and pre-pandemic uninfected donors were collected, heat-inactivated for 1h at 56°C and stored at −80°C until ready to use in subsequent experiments. Plasma from uninfected donors were used as negative controls and used to calculate the seropositivity threshold in our ELISA, cell-based ELISA, and flow cytometry assays. The RBD-specific monoclonal antibody CR302244 (link) was used as a positive control in our ELISAs, cell-based ELISAs, and flow cytometry assays and was previously described14 ,16 (link),24 (link). Horseradish peroxidase (HRP)-conjugated antibodies able to detect all Ig isotypes (anti-human IgM+IgG+IgA; Jackson ImmunoResearch Laboratories, Inc.) or specific for the Fc region of human IgG (Invitrogen), the Fc region of human IgM (Jackson ImmunoResearch Laboratories, inc.) or the Fc region of human IgA (Jackson ImmunoResearch Laboratories, inc) were used as secondary antibodies to detect antibody binding in ELISA and cell-based ELISA experiments. Alexa Fluor-647-conjugated goat anti-human Abs able to detect all Ig isotypes (anti-human IgM+IgG+IgA; Jackson ImmunoResearch Laboratories, Inc.) were used as secondary antibodies to detect plasma binding in flow cytometry experiments.
Anand S.P., Prévost J., Nayrac M., Beaudoin-Bussières G., Benlarbi M., Gasser R., Brassard N., Laumaea A., Gong S.Y., Bourassa C., Brunet-Ratnasingham E., Medjahed H., Gendron-Lepage G., Goyette G., Gokool L., Morrisseau C., Bégin P., Martel-Laferrière V., Tremblay C., Richard J., Bazin R., Duerr R., Kaufmann D.E, & Finzi A. (2021). Longitudinal analysis of humoral immunity against SARS-CoV-2 Spike in convalescent individuals up to 8 months post-symptom onset. Cell Reports Medicine, 2(6), 100290.
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3

Characterization of SARS-CoV-2 Antibodies

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Plasma from SARS-CoV-2 naive and PI donors were collected, heat-inactivated for 1 h at 56°C and stored at −80°C until ready to use in subsequent experiments. Plasma from uninfected donors collected before the pandemic were used as negative controls and used to calculate the seropositivity threshold in our ELISA, cell-based ELISA, ADCC and flow cytometry assays (see below). The RBD-specific monoclonal antibody CR3022 was used as a positive control in our ELISA, cell-based ELISA, and flow cytometry assays and was previously described (Anand et al., 2021a (link); Beaudoin-Bussières et al., 2020 (link); Prévost et al., 2020 (link)). Horseradish peroxidase (HRP)-conjugated Abs able to detect all Ig isotypes (anti-human IgM+IgG+IgA; Jackson ImmunoResearch Laboratories) or specific for the Fc region of human IgG (Invitrogen), the Fc region of human IgM (Jackson ImmunoResearch Laboratories) or the Fc region of human IgA (Jackson ImmunoResearch Laboratories) were used as secondary Abs to detect Ab binding in ELISA and cell-based ELISA experiments. Alexa Fluor-647-conjugated goat anti-human Abs able to detect all Ig isotypes (anti-human IgM+IgG+IgA; Jackson ImmunoResearch Laboratories) were used as secondary Ab to detect plasma binding in flow cytometry experiments.
Tauzin A., Nayrac M., Benlarbi M., Gong S.Y., Gasser R., Beaudoin-Bussières G., Brassard N., Laumaea A., Vézina D., Prévost J., Anand S.P., Bourassa C., Gendron-Lepage G., Medjahed H., Goyette G., Niessl J., Tastet O., Gokool L., Morrisseau C., Arlotto P., Stamatatos L., McGuire A.T., Larochelle C., Uchil P., Lu M., Mothes W., De Serres G., Moreira S., Roger M., Richard J., Martel-Laferrière V., Duerr R., Tremblay C., Kaufmann D.E, & Finzi A. (2021). A single dose of the SARS-CoV-2 vaccine BNT162b2 elicits Fc-mediated antibody effector functions and T cell responses. Cell Host & Microbe, 29(7), 1137-1150.e6.
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4

Isolation and Activation of Human B Cells

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PBMC were prepared by Ficoll-Paque density centrifugation. Isolated B cells were prepared from PBMC using the Dynabeads Untouched Human B Cells Kit (11351D, Thermo Fisher Scientific) according to the manufacturer’s recommended protocol. Purity of the B cell suspension was assessed by flow cytometry, indicating a 99% CD19+ cell population consistent with high purity of B cells. B cells were resuspended 0.5 million/ml ml in RPMI 1640 containing 10% heat-inactivated human serum, 2 mM l-glutamine, 1 mM Na-pyruvate, 0.1 mM nonessential amino acids, 50 μM β-mercaptoethanol, streptomycin (100 μg/ml), and penicillin (100 U/ml), alone, or supplemented with IL-4 (20 ng/ml; PeproTech, Rocky Hill, NJ) plus anti-human IgA + IgG + IgM (30 μg/ml; Jackson ImmunoResearch Laboratories, West Grove, PA) plus sCD40L (1000 ng/ml; Enzo Life Sciences, Farmingdale, NY). B cells (0.1 million) were added in 0.2 ml per well in 96-well U-bottom tissue culture plates and incubated at 37°C in 5% CO2. After 24 hours, supernatants were harvested and frozen until evaluation.
Goel G., Tye-Din J.A., Qiao S.W., Russell A.K., Mayassi T., Ciszewski C., Sarna V.K., Wang S., Goldstein K.E., Dzuris J.L., Williams L.J., Xavier R.J., Lundin K.E., Jabri B., Sollid L.M, & Anderson R.P. (2019). Cytokine release and gastrointestinal symptoms after gluten challenge in celiac disease. Science Advances, 5(8), eaaw7756.
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5

Quantifying T and B Cell Activation

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Frozen PBMCs were thawed, plated, and rested in complete RPMI at 37 °C prior to stimulation with 5 µg/mL anti-CD3 (UCHT1) and 5 µg/mL anti-CD28 (CD28.2) antibodies (both from eBioscience) or anti-human IgA/IgG/IgM (Jackson ImmunoResearch, West Grove, PA) for the indicated durations. Cells were fixed in 2% paraformaldehyde and permeabilized with 100% methanol prior to staining for CD14 (M5E5, BioLegend), CD4 (RPA-T4, eBioscience), CD8 (SK1, eBioscience), CD19 (HIB19, eBioscience), and phospho-Erk (Thr202/Tyr204, MILAN8R, eBioscience). Cells were acquired on a BD FACSCanto flow cytometer (BD Biosciences). After exclusion of doublets and monocytes via CD14, T cells were gated as CD19-, and B cells were gated as CD4-CD8-CD19+ cells (Supplementary Fig. 1B). The fold change in the median fluorescence intensity was calculated based on the whole unstimulated population of the respective sample.
Neumann J., Van Nieuwenhove E., Terry L.E., Staels F., Knebel T.R., Welkenhuyzen K., Ahmadzadeh K., Baker M.R., Gerbaux M., Willemsen M., Barber J.S., Serysheva I.I., De Waele L., Vermeulen F., Schlenner S., Meyts I., Yule D.I., Bultynck G., Schrijvers R., Humblet-Baron S, & Liston A. (2022). Disrupted Ca2+ homeostasis and immunodeficiency in patients with functional IP3 receptor subtype 3 defects. Cellular and Molecular Immunology, 20(1), 11-25.
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6

Functional analysis of immune cells

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Frozen PBMCs were thawed, plated and rested in complete RPMI at 37 °C prior to stimulation with 5 µg/mL aCD3 (UCHT1) and 5 µg/mL aCD28 (CD28.2, both eBioscience) or anti-human IgA/IgG/IgM (Jackson ImmunoResearch, West Grove, PA), for durations as indicated. Cells were fixed in 2 % paraformaldehyde and permeabilized with 100 % methanol, prior to staining for CD14 (M5E5, BioLegend), CD4 (RPA-T4, eBioscience), CD8 (SK1, eBioscience), CD19 (HIB19, eBioscience), phospho-Erk (Thr202/Tyr204, MILAN8R, eBioscience) and phospho-PLCγ1 (Tyr783, both Biolegend). Cells were acquired on a BD FACSCanto (BD Biosciences). After exclusion of doublets and monocytes via CD14, T cells were gated as being CD19 -and B cells as CD19 + . The fold change of the median fluorescence intensity was calculated based on the average of all healthy controls within one repeat of the experiment.
Neumann J., Nieuwenhove E.V., Terry L.E., Staels F., Knebel T.R., Welkenhuyzen K., Baker M.R., Gerbaux M., Willemsen M., Barber J.S., Serysheva I.I., Waele L.D., Vermeulen F., Meyts I., Yule D.I., Bultynck G., Schrijvers R., Humblet‐Baron S., & Liston A. (2021). Disrupted Ca2+ homeostasis and immunodeficiency in patients with functional Inositol 1,4,5-trisphosphate receptor subtype 3 defects.
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7

SARS-CoV-2 Antibody Detection Protocol

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Plasmas were isolated by Ficoll density gradient from blood samples. Plasmas were heat-inactivated for 1 h at 56 °C and stored at −80 °C until use. We used plasma samples from uninfected/unvaccinated donors collected before the pandemic as negative controls in ELISA, ADCC, and cytometry assays, and to calculate the seropositivity threshold. The CR3022 monoclonal antibody (mAb) (a receptor-binding domain (RBD)-specific monoclonal antibody) and the CV3-25 mAb (a conformationally independent S2-specific mAb) were used as positive controls in ELISA using the ancestral RBD and flow cytometry assays, respectively [10 (link),30 (link),31 (link),32 (link)]. We used as secondary antibodies (Abs) a Horseradish peroxidase (HRP)-conjugated Abs that detect the Fc region of human IgG (Invitrogen) in ELISA assays and Alexa Fluor-647-conjugated goat antihuman antibodies detecting all Ig isotypes (antihuman IgM, IgG, IgA; Jackson ImmunoResearch Laboratories, cat # 109-605-064) in flow cytometry experiments.
Tauzin A., Beaudoin-Bussières G., Benlarbi M., Nayrac M., Bo Y., Gendron-Lepage G., Medjahed H., Perreault J., Gokool L., Arlotto P., Morrisseau C., Tremblay C., Kaufmann D.E., Martel-Laferrière V., Levade I., Côté M., Bazin R, & Finzi A. (2023). Humoral Responses Elicited after a Fifth Dose of SARS-CoV-2 mRNA Bivalent Vaccine. Viruses, 15(9), 1926.
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8

SARS-CoV-2 Serological Assays using Plasma Samples

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Plasma from SOTR and HCW were collected, heat-inactivated for 1 hour at 56°C and stored at −80°C until ready to use in subsequent experiments. Plasma from uninfected donors collected before the pandemic were used as negative controls and used to calculate the seropositivity threshold in our ELISA, ADCC and flow cytometry assays (see below). The RBD-specific monoclonal antibody CR3022 was used as a positive control in our ELISA, and the CV3-25 antibody in flow cytometry assays and were previously described (Anand et al., 2020 ; Beaudoin-Bussières et al., 2020 (link); Jennewein et al., 2021 (link); Meulen et al., 2006 (link); Prévost et al., 2020 (link)). Horseradish peroxidase (HRP)-conjugated Abs able to recognize the Fc region of human IgG (Invitrogen) were used as secondary Abs in ELISA experiments. Alexa Fluor-647-conjugated goat anti-human Abs able to detect all Ig isotypes (anti-human IgM+IgG+IgA; Jackson ImmunoResearch Laboratories) were used as secondary Abs to detect plasma binding in flow cytometry experiments.
Tauzin A., Beaudoin-Bussières G., Gong S.Y., Chatterjee D., Gendron-Lepage G., Bourassa C., Goyette G., Racine N., Khrifi Z., Turgeon J., Tremblay C., Martel-Laferrière V., Kaufmann D.E., Cardinal H., Cloutier M., Bazin R., Duerr R., Dieudé M., Hébert M.J, & Finzi A. (2022). Humoral immune responses against SARS-CoV-2 Spike variants after mRNA vaccination in solid organ transplant recipients. iScience, 25(9), 104990.
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9

SARS-CoV-2 Antibody Characterization Protocol

Cited 2 times
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Plasma from SARS-CoV-2 naïve and PI donors were collected, heat-inactivated for 1 h at 56°C and stored at −80°C until ready to use in subsequent experiments. Plasma from uninfected donors collected before the pandemic were used as negative controls and used to calculate the seropositivity threshold in our ELISA, ADCC and flow cytometry assays (see below). The RBD-specific monoclonal antibody CR3022 was used as a positive control in ELISA assays, and the CV3-25 antibody in flow cytometry assays and were previously described (Anand et al., 2020 ; Beaudoin-Bussières et al., 2020 (link); Jennewein et al., 2021 (link); Meulen et al., 2006 (link); Prévost et al., 2020 (link)). Horseradish peroxidase (HRP)-conjugated Abs able to detect the Fc region of human IgG (Invitrogen) was used as secondary Abs to detect Ab binding in ELISA experiments. Alexa Fluor-647-conjugated goat anti-human Abs able to detect all Ig isotypes (anti-human IgM+IgG+IgA; Jackson ImmunoResearch Laboratories) were used as secondary Ab to detect plasma binding in flow cytometry experiments.
Tauzin A., Gong S.Y., Chatterjee D., Ding S., Painter M.M., Goel R.R., Beaudoin-Bussières G., Marchitto L., Boutin M., Laumaea A., Okeny J., Gendron-Lepage G., Bourassa C., Medjahed H., Goyette G., Williams J.C., Bo Y., Gokool L., Morrisseau C., Arlotto P., Bazin R., Fafard J., Tremblay C., Kaufmann D.E., De Serres G., Richard J., Côté M., Duerr R., Martel-Laferrière V., Greenplate A.R., Wherry E.J, & Finzi A. (2022). A boost with SARS-CoV-2 BNT162b2 mRNA vaccine elicits strong humoral responses independently of the interval between the first two doses. Cell Reports, 41(4), 111554.
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10

SARS-CoV-2 Spike Protein Detection

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Plasma samples were either recovered from whole blood or directly obtained from the PlasCov biobank [25 (link)], heat-inactivated for 1 h at 56 °C and stored at −80 °C until use in subsequent experiments. Pre-pandemic plasma samples were used as negative controls in cytometry assays (data not shown). The conformationally independent S2-specific monoclonal antibody CV3-25 was used as a positive control and to normalize Spike expression in flow cytometry assays, as described [4 (link),26 (link),27 (link),28 (link),29 (link)]. Alexa Fluor-647-conjugated goat anti-human antibodies (Abs) able to detect all Ig isotypes (anti-human IgM, IgG, IgA; Jackson ImmunoResearch Laboratories, Cat # 109-605-064) were used as secondary Abs to detect plasma binding in flow cytometry experiments.
Tauzin A., Benlarbi M., Medjahed H., Grégoire Y., Perreault J., Gendron-Lepage G., Gokool L., Morrisseau C., Arlotto P., Tremblay C., Kaufmann D.E., Martel-Laferrière V., Levade I., Côté M., De Serres G., Bazin R, & Finzi A. (2023). Humoral Responses against BQ.1.1 Elicited after Breakthrough Infection and SARS-CoV-2 mRNA Vaccination. Vaccines, 11(2), 242.
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