igm antibody  (Thermo Fisher)

Journal: Nature Biomedical Engineering

Article Title: Ultrasensitive high-resolution profiling of early seroconversion in patients with COVID-19

doi: 10.1038/s41551-020-00611-x

Figure Lengend Snippet: Profiling seroconversion in COVID-19. Simoa serological assay results for IgG, IgM and IgA against the four viral targets (spike, S1 subunit, RBD and nucleocapsid) for pre-pandemic samples (light blue; n = 200), nasopharyngeal (NP) PCR-negative samples (dark blue; n = 100) and SARS-CoV-2-positive samples (black (immunocompetent; n = 141) or red (immunosuppressed; n = 31)). The SARS-CoV-2-positive samples were divided into four groups according to time since since the first positive nasopharyngeal RT–PCR test. Black lines indicate the median normalized AEB value of each population.

Article Snippet: Anti-human immunoglobulin antibodies were diluted in Homebrew Detector/Sample Diluent to final concentrations of 7.73 ng ml−1 IgG (Bethyl Labratories; A80-148B), 216 ng ml−1 IgM (Thermo Fisher Scientific; MII0401) and 150 ng ml−1 IgA (Abcam; ab214003).

Techniques: Serologic Assay, Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction

Journal: Nature Biomedical Engineering

Article Title: Ultrasensitive high-resolution profiling of early seroconversion in patients with COVID-19

doi: 10.1038/s41551-020-00611-x

Figure Lengend Snippet: Classification of COVID-19 using Simoa serological assays. a , Left: ROC curves for all positive cases ( n = 141) in the pre-pandemic cohort ( n = 199) using either the full marker panel model (green), the early-stage model (blue) or the late-stage model (red). Middle: ROC curves for late-stage cases (more than 1 week after positive nasopharyngeal PCR test; n = 50) in the pre-pandemic cohort ( n = 199) using either the full marker panel model (green) or the late-stage model (red). Right: ROC curves for early-stage cases (first week after positive nasopharyngeal PCR test; n = 91) in the pre-pandemic cohort ( n = 199) using either the full marker panel model (green) or the early-stage model (blue). AUC values and 95% confidence intervals (CIs) are shown for each graph. b , Simoa serological assay results for IgG, IgM and IgA against the four viral targets (spike, S1 subunit, RBD and nucleocapsid) for pre-pandemic samples (light blue; n = 232) and nasopharyngeal RT–PCR-positive samples (grey; n = 68). Statistical significance was determined using Mann–Whitney U -tests. All U -tests were two tailed and did not correct for multiple comparisons. Black lines indicate the median normalized AEB value of each population.

Article Snippet: Anti-human immunoglobulin antibodies were diluted in Homebrew Detector/Sample Diluent to final concentrations of 7.73 ng ml−1 IgG (Bethyl Labratories; A80-148B), 216 ng ml−1 IgM (Thermo Fisher Scientific; MII0401) and 150 ng ml−1 IgA (Abcam; ab214003).

Techniques: Marker, Polymerase Chain Reaction, Serologic Assay, Reverse Transcription Polymerase Chain Reaction, MANN-WHITNEY, Two Tailed Test

Journal: Nature Biomedical Engineering

Article Title: Ultrasensitive high-resolution profiling of early seroconversion in patients with COVID-19

doi: 10.1038/s41551-020-00611-x

Figure Lengend Snippet: Schematic of the Simoa serological assay. Plasma is incubated with four types of dye-encoded beads that are each coupled to one of four viral targets (spike, S1, RBD and nucleocapsid). IgG, IgA and IgM antibodies specific to the SARS-CoV-2 targets bind to the viral antigen-coated beads. After washing, beads are introduced to biotinylated anti-human immunoglobulin antibodies to label either IgG, IgM or IgA in the different reactions. After additional washes, the enzyme SβG is introduced. The beads are washed, resuspended in fluorogenic RGP and loaded into a 216,000-microwell array for multicolour imaging.

Article Snippet: Anti-human immunoglobulin antibodies were diluted in Homebrew Detector/Sample Diluent to final concentrations of 7.73 ng ml−1 IgG (Bethyl Labratories; A80-148B), 216 ng ml−1 IgM (Thermo Fisher Scientific; MII0401) and 150 ng ml−1 IgA (Abcam; ab214003).

Techniques: Serologic Assay, Incubation, Imaging

Journal: Infection and Immunity

Article Title: Major Histocompatibility Complex Class II-Restricted, CD4+ T Cell-Dependent and -Independent Mechanisms Are Required for Vaccine-Induced Protective Immunity against Coxiella burnetii

doi: 10.1128/IAI.00824-19

Figure Lengend Snippet: MHC-I and MHC-II are involved in antibody isotype switching. B2m KO, MHC-II KO, and WT C57BL/6 mice were vaccinated s.c. with 10 μg of PIV and challenged i.p. with 1 × 10 7 genomic copies of C. burnetii NMI 28 dpv. Mice receiving Alhydrogel adjuvant alone served as unvaccinated controls. C. burnetii NMI-specific serum IgM (A to C) and IgG (D to F) were evaluated weekly following vaccination. Specific IgM (G) and IgG (H) were also evaluated 14 dpi. Each experimental group includes five mice, with error bars representing the standard deviations from the mean. **, P

Article Snippet: Plates were washed with PBS-T buffer and then incubated with 100 μl of diluted horseradish peroxidase-conjugated goat anti-mouse IgM or IgG (1:4,000 to 1:8,000) at room temperature for 1 h. The plates were again washed with PBS-T, followed by the addition of 100 μl of TMB substrate (Thermo Fisher Scientific, Waltham, MA).

Techniques: Mouse Assay

Journal: Infection and Immunity

Article Title: Major Histocompatibility Complex Class II-Restricted, CD4+ T Cell-Dependent and -Independent Mechanisms Are Required for Vaccine-Induced Protective Immunity against Coxiella burnetii

doi: 10.1128/IAI.00824-19

Figure Lengend Snippet: CD4 + T cells are important for antibody isotype switching. WT, CD4 KO, and MHC-II KO mice were vaccinated and challenged as previously described. Mice receiving Alhydrogel adjuvant alone served as unvaccinated controls. C. burnetii NMI-specific serum IgM and IgG were evaluated weekly until 28 dpv (A) and 14 dpi (B). Each experimental group includes five mice, with error bars representing the standard deviations from the mean. *, P

Article Snippet: Plates were washed with PBS-T buffer and then incubated with 100 μl of diluted horseradish peroxidase-conjugated goat anti-mouse IgM or IgG (1:4,000 to 1:8,000) at room temperature for 1 h. The plates were again washed with PBS-T, followed by the addition of 100 μl of TMB substrate (Thermo Fisher Scientific, Waltham, MA).

Techniques: Mouse Assay

Journal: Scientific Reports

Article Title: Evidence that endogenous formaldehyde produces immunogenic and atherogenic adduct epitopes

doi: 10.1038/s41598-017-11289-8

Figure Lengend Snippet: The immunogenicity of M2FA-lysine-BSA in the absence of adjuvant and anti-M2FA antibody titers in intact mice. C57BL/6 mice were injected i.p. with M2FA-lysine-BSA or BSA in the absence of adjuvant. The antibody titers of IgG ( a ) and IgM ( b ) against M2FA-lysine were detected using MFA-6ACA-KLH-coated plates. The anti-M2AA antibody titers were clearly increased in M2FA-lysine-BSA-immunized mice compared to the controls (BSA-treated mice). Values are mean and SD. ( c ) Intact female C57BL/6 mice (n = 4 or 5 per group) with different ages showed significantly different anti-M2FA IgG titers. Values are mean and SD. (*p

Article Snippet: Malondialdehyde bis(dimethyl) acetal, Dynabeads M-270 Amine, CarboxyLink Kit, the Imject EDC mcKLH Spin Kit, goat anti-rabbit IgG (H + L) antibody with HRP, goat anti-human IgG/IgM secondary antibody, and goat anti-mouse IgM secondary antibody were obtained from Thermo Scientific (Rockford, IL).

Techniques: Mouse Assay, Injection

Journal: Scientific Reports

Article Title: Evidence that endogenous formaldehyde produces immunogenic and atherogenic adduct epitopes

doi: 10.1038/s41598-017-11289-8

Figure Lengend Snippet: Serum anti-M2FA IgG and IgM antibody levels in wild-type and ApoE −/− mice and immunohistochemical detection of M2FA-epitopes in heart valve of ApoE −/− mice. The anti-M2FA IgG ( a ) and IgM ( b ) antibody levels showed significant differences between wild-type and ApoE −/− mice with the M2FA-6ACA-BSA ELISAs (**p

Article Snippet: Malondialdehyde bis(dimethyl) acetal, Dynabeads M-270 Amine, CarboxyLink Kit, the Imject EDC mcKLH Spin Kit, goat anti-rabbit IgG (H + L) antibody with HRP, goat anti-human IgG/IgM secondary antibody, and goat anti-mouse IgM secondary antibody were obtained from Thermo Scientific (Rockford, IL).

Techniques: Mouse Assay, Immunohistochemistry