igm antibodies Search Results


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  • 99
    Thermo Fisher igm
    Tet2 deletion in hematopoietic cells induces GC hyperplasia. A, Representative flow cytometry plot and quantification of <t>(B220</t> + CD95 + GL7 + ) GC B-cells from Vav-Cre/ Tet2 +/+ (n=5) and Vav-Cre/ Tet2 −/− (n=5) mice at day 10 after immunization with SRBC. B , Quantification of flow cytometry data corresponding to total B cells (B220 + ), mature B cells (B220 + IgD + <t>IgM</t> + ), transitional B cells (B220 + IgD int IgM + ), follicular B cells (B220 + CD23 + CD21 + ), marginal zone B cells (B220 + CD23 low CD21 + ) and plasmablasts/PC (CD138 + ) in the spleens of Vav-Cre/ Tet2 +/+ and Vav-Cre/ Tet2 −/− mice. C , Representative histologic sections of formalin-fixed, paraffin-embedded spleens from Vav-Cre/ Tet2 +/+ and Vav-Cre/ Tet2 −/− mice. Sections were stained with H E and antibodies specific for B220, PNA and Ki-67. D , E , Quantification of GC area ( D ) and number of GCs ( E ) in the spleens of Vav-Cre/ Tet2 +/+ and Vav-Cre/ Tet2 −/− mice. two-tailed t test, ***p
    Igm, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 4205 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/igm/product/Thermo Fisher
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    igm - by Bioz Stars, 2020-11
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    99
    Millipore polyclonal antibodies
    Tet2 deletion in hematopoietic cells induces GC hyperplasia. A, Representative flow cytometry plot and quantification of <t>(B220</t> + CD95 + GL7 + ) GC B-cells from Vav-Cre/ Tet2 +/+ (n=5) and Vav-Cre/ Tet2 −/− (n=5) mice at day 10 after immunization with SRBC. B , Quantification of flow cytometry data corresponding to total B cells (B220 + ), mature B cells (B220 + IgD + <t>IgM</t> + ), transitional B cells (B220 + IgD int IgM + ), follicular B cells (B220 + CD23 + CD21 + ), marginal zone B cells (B220 + CD23 low CD21 + ) and plasmablasts/PC (CD138 + ) in the spleens of Vav-Cre/ Tet2 +/+ and Vav-Cre/ Tet2 −/− mice. C , Representative histologic sections of formalin-fixed, paraffin-embedded spleens from Vav-Cre/ Tet2 +/+ and Vav-Cre/ Tet2 −/− mice. Sections were stained with H E and antibodies specific for B220, PNA and Ki-67. D , E , Quantification of GC area ( D ) and number of GCs ( E ) in the spleens of Vav-Cre/ Tet2 +/+ and Vav-Cre/ Tet2 −/− mice. two-tailed t test, ***p
    Polyclonal Antibodies, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 3662 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/polyclonal antibodies/product/Millipore
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    99
    Thermo Fisher goat anti mouse igm heavy chain secondary antibody
    Tet2 deletion in hematopoietic cells induces GC hyperplasia. A, Representative flow cytometry plot and quantification of <t>(B220</t> + CD95 + GL7 + ) GC B-cells from Vav-Cre/ Tet2 +/+ (n=5) and Vav-Cre/ Tet2 −/− (n=5) mice at day 10 after immunization with SRBC. B , Quantification of flow cytometry data corresponding to total B cells (B220 + ), mature B cells (B220 + IgD + <t>IgM</t> + ), transitional B cells (B220 + IgD int IgM + ), follicular B cells (B220 + CD23 + CD21 + ), marginal zone B cells (B220 + CD23 low CD21 + ) and plasmablasts/PC (CD138 + ) in the spleens of Vav-Cre/ Tet2 +/+ and Vav-Cre/ Tet2 −/− mice. C , Representative histologic sections of formalin-fixed, paraffin-embedded spleens from Vav-Cre/ Tet2 +/+ and Vav-Cre/ Tet2 −/− mice. Sections were stained with H E and antibodies specific for B220, PNA and Ki-67. D , E , Quantification of GC area ( D ) and number of GCs ( E ) in the spleens of Vav-Cre/ Tet2 +/+ and Vav-Cre/ Tet2 −/− mice. two-tailed t test, ***p
    Goat Anti Mouse Igm Heavy Chain Secondary Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 272 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Becton Dickinson anti igm
    Impaired release of immature bone marrow B cells by CXCR4 antagonism in the absence of S1P1 receptor. (A) Mice were injected with the CXCR4 antagonist AMD3100 or vehicle alone, and bone marrow and blood were collected after 90 min. Immature bone marrow B cells in the blood (B220 + CD93 + ) were gated as <t>IgM</t> + <t>IgD</t> − and IgM + IgD low , and quantified from control and B- S1pr1 KO mice. Results are shown as absolute numbers in 250 µl of blood. Bars represent mean values, and the closed circles are individual mice. Data are representative of three experiments. *, P
    Anti Igm, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 94/100, based on 991 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore anti human igm mu chain specific f ab 2 fragment peroxidase antibody
    Impaired release of immature bone marrow B cells by CXCR4 antagonism in the absence of S1P1 receptor. (A) Mice were injected with the CXCR4 antagonist AMD3100 or vehicle alone, and bone marrow and blood were collected after 90 min. Immature bone marrow B cells in the blood (B220 + CD93 + ) were gated as <t>IgM</t> + <t>IgD</t> − and IgM + IgD low , and quantified from control and B- S1pr1 KO mice. Results are shown as absolute numbers in 250 µl of blood. Bars represent mean values, and the closed circles are individual mice. Data are representative of three experiments. *, P
    Anti Human Igm Mu Chain Specific F Ab 2 Fragment Peroxidase Antibody, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    SouthernBiotech anti igm
    SVP[Rapa] treatment enables AAV8 re-administration in nonhuman primates. a Protocol outline. Male naive cynomolgus monkeys were treated i.v. with 2 × 10 12 vg kg −1 of AAV8-Gaa vector and either SVP[Rapa] (3 mg kg −1 , n = 2, SVP[Rapa]#1 and SVP[Rapa]#2) or SVP[empty] control ( n = 1) and then challenged i.v. on day 30 with 2 × 10 12 vg kg − 1 of AAV8-hF.IX vector and either SVP[Rapa] or SVP[empty] control, as described above. b , c Analysis of b anti-AAV8 <t>IgG</t> antibodies and c anti-AAV8 <t>IgM</t> antibody responses measured by ELISA. d Analysis of anti-AAV8 neutralizing antibodies (NAb) measured with a cell-based neutralization assay. e Analysis of anti-AAV8 IgG secreting B cells in splenocytes measured by B ELISpot. Data are shown as individual replicates and the bars represent mean ± s.d. The dotted line indicates the threshold for positivity corresponding to 50 spot forming units (SFU) per million cells. f Plasma hF.IX antigen levels quantified by ELISA at the indicated time points following administration of AAV8-hF.IX vector. g AAV8-hF.IX vector genome copy number (VGCN) per diploid genome in liver. The symbols represent individual liver lobes (left, right, caudate and quadrate) and the bars represent the mean ± s.d. (4 liver lobes per monkey; one-way ANOVA with Tukey post hoc test, ** p
    Anti Igm, supplied by SouthernBiotech, used in various techniques. Bioz Stars score: 95/100, based on 610 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Novus Biologicals rabbit polyclonal antibody
    Co-expression of sortilin, APLP2, or both with PCSK9 has no major effect on LDLR degradation. Huh7 cells were transfected with a total of 3 μg using 1 μg of each vector encoding for either a control protein 7B2 (−), sortilin (+), APLP2 (+), or PCSK9 (+), as indicated. After 48 h, lysates were analyzed by Western blotting for expression of the LDLR, sortilin-Myc, APLP2-V5, intracellular pro- and mature-PCSK9-V5, and β-actin. Media were analyzed for secreted endogenous and overexpressed PCSK9-V5 using a rabbit <t>polyclonal</t> human PCSK9 antibody. Quantification of LDLR expression was normalized against that of β-actin. These data are representative of at least 3 different experiments showing similar results.
    Rabbit Polyclonal Antibody, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 99/100, based on 944 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher goat anti mouse igm heavy chain cross adsorbed secondary antibody
    Co-expression of sortilin, APLP2, or both with PCSK9 has no major effect on LDLR degradation. Huh7 cells were transfected with a total of 3 μg using 1 μg of each vector encoding for either a control protein 7B2 (−), sortilin (+), APLP2 (+), or PCSK9 (+), as indicated. After 48 h, lysates were analyzed by Western blotting for expression of the LDLR, sortilin-Myc, APLP2-V5, intracellular pro- and mature-PCSK9-V5, and β-actin. Media were analyzed for secreted endogenous and overexpressed PCSK9-V5 using a rabbit <t>polyclonal</t> human PCSK9 antibody. Quantification of LDLR expression was normalized against that of β-actin. These data are representative of at least 3 different experiments showing similar results.
    Goat Anti Mouse Igm Heavy Chain Cross Adsorbed Secondary Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 534 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Thermo Fisher f ab 2 goat anti human igg iga igm secondary antibody
    Co-expression of sortilin, APLP2, or both with PCSK9 has no major effect on LDLR degradation. Huh7 cells were transfected with a total of 3 μg using 1 μg of each vector encoding for either a control protein 7B2 (−), sortilin (+), APLP2 (+), or PCSK9 (+), as indicated. After 48 h, lysates were analyzed by Western blotting for expression of the LDLR, sortilin-Myc, APLP2-V5, intracellular pro- and mature-PCSK9-V5, and β-actin. Media were analyzed for secreted endogenous and overexpressed PCSK9-V5 using a rabbit <t>polyclonal</t> human PCSK9 antibody. Quantification of LDLR expression was normalized against that of β-actin. These data are representative of at least 3 different experiments showing similar results.
    F Ab 2 Goat Anti Human Igg Iga Igm Secondary Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher peroxidase conjugated anti rabbit
    Co-expression of sortilin, APLP2, or both with PCSK9 has no major effect on LDLR degradation. Huh7 cells were transfected with a total of 3 μg using 1 μg of each vector encoding for either a control protein 7B2 (−), sortilin (+), APLP2 (+), or PCSK9 (+), as indicated. After 48 h, lysates were analyzed by Western blotting for expression of the LDLR, sortilin-Myc, APLP2-V5, intracellular pro- and mature-PCSK9-V5, and β-actin. Media were analyzed for secreted endogenous and overexpressed PCSK9-V5 using a rabbit <t>polyclonal</t> human PCSK9 antibody. Quantification of LDLR expression was normalized against that of β-actin. These data are representative of at least 3 different experiments showing similar results.
    Peroxidase Conjugated Anti Rabbit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 547 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Bio-Rad goat anti rabbit secondary ab
    Co-expression of sortilin, APLP2, or both with PCSK9 has no major effect on LDLR degradation. Huh7 cells were transfected with a total of 3 μg using 1 μg of each vector encoding for either a control protein 7B2 (−), sortilin (+), APLP2 (+), or PCSK9 (+), as indicated. After 48 h, lysates were analyzed by Western blotting for expression of the LDLR, sortilin-Myc, APLP2-V5, intracellular pro- and mature-PCSK9-V5, and β-actin. Media were analyzed for secreted endogenous and overexpressed PCSK9-V5 using a rabbit <t>polyclonal</t> human PCSK9 antibody. Quantification of LDLR expression was normalized against that of β-actin. These data are representative of at least 3 different experiments showing similar results.
    Goat Anti Rabbit Secondary Ab, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore isotype control antibody
    Co-expression of sortilin, APLP2, or both with PCSK9 has no major effect on LDLR degradation. Huh7 cells were transfected with a total of 3 μg using 1 μg of each vector encoding for either a control protein 7B2 (−), sortilin (+), APLP2 (+), or PCSK9 (+), as indicated. After 48 h, lysates were analyzed by Western blotting for expression of the LDLR, sortilin-Myc, APLP2-V5, intracellular pro- and mature-PCSK9-V5, and β-actin. Media were analyzed for secreted endogenous and overexpressed PCSK9-V5 using a rabbit <t>polyclonal</t> human PCSK9 antibody. Quantification of LDLR expression was normalized against that of β-actin. These data are representative of at least 3 different experiments showing similar results.
    Isotype Control Antibody, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 571 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher igm antibody
    Co-localization of PAB-reactive PLTA antigen, <t>IgA,</t> <t>IgM,</t> and C3c detected by double fluorescence immunohistochemistry. A: Anti-IgA antibody (red) vs PAB antibody (green) after MT treatment, B: anti-IgM antibody (red) vs PAB antibody (green) after MT treatment, C: anti-IgM antibody (red) vs anti-IgA antibody (green), and D: anti-C3c antibody (red) vs PAB antibody (green) after MT treatment. Many PAB-reactive SRBs were also positive for IgA, IgM, and C3c, showing yellow-colored double-positive signals (A, B, and D, respectively). Both IgA and IgM colocalized with these SRBs, indicated by yellow-colored double-positive signals (C).
    Igm Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 104 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/igm antibody/product/Thermo Fisher
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    R&D Systems goat polyclonal antibody
    Co-localization of PAB-reactive PLTA antigen, <t>IgA,</t> <t>IgM,</t> and C3c detected by double fluorescence immunohistochemistry. A: Anti-IgA antibody (red) vs PAB antibody (green) after MT treatment, B: anti-IgM antibody (red) vs PAB antibody (green) after MT treatment, C: anti-IgM antibody (red) vs anti-IgA antibody (green), and D: anti-C3c antibody (red) vs PAB antibody (green) after MT treatment. Many PAB-reactive SRBs were also positive for IgA, IgM, and C3c, showing yellow-colored double-positive signals (A, B, and D, respectively). Both IgA and IgM colocalized with these SRBs, indicated by yellow-colored double-positive signals (C).
    Goat Polyclonal Antibody, supplied by R&D Systems, used in various techniques. Bioz Stars score: 99/100, based on 770 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Bio-Rad goat anti rabbit antibody
    Co-localization of PAB-reactive PLTA antigen, <t>IgA,</t> <t>IgM,</t> and C3c detected by double fluorescence immunohistochemistry. A: Anti-IgA antibody (red) vs PAB antibody (green) after MT treatment, B: anti-IgM antibody (red) vs PAB antibody (green) after MT treatment, C: anti-IgM antibody (red) vs anti-IgA antibody (green), and D: anti-C3c antibody (red) vs PAB antibody (green) after MT treatment. Many PAB-reactive SRBs were also positive for IgA, IgM, and C3c, showing yellow-colored double-positive signals (A, B, and D, respectively). Both IgA and IgM colocalized with these SRBs, indicated by yellow-colored double-positive signals (C).
    Goat Anti Rabbit Antibody, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 95/100, based on 359 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Tet2 deletion in hematopoietic cells induces GC hyperplasia. A, Representative flow cytometry plot and quantification of (B220 + CD95 + GL7 + ) GC B-cells from Vav-Cre/ Tet2 +/+ (n=5) and Vav-Cre/ Tet2 −/− (n=5) mice at day 10 after immunization with SRBC. B , Quantification of flow cytometry data corresponding to total B cells (B220 + ), mature B cells (B220 + IgD + IgM + ), transitional B cells (B220 + IgD int IgM + ), follicular B cells (B220 + CD23 + CD21 + ), marginal zone B cells (B220 + CD23 low CD21 + ) and plasmablasts/PC (CD138 + ) in the spleens of Vav-Cre/ Tet2 +/+ and Vav-Cre/ Tet2 −/− mice. C , Representative histologic sections of formalin-fixed, paraffin-embedded spleens from Vav-Cre/ Tet2 +/+ and Vav-Cre/ Tet2 −/− mice. Sections were stained with H E and antibodies specific for B220, PNA and Ki-67. D , E , Quantification of GC area ( D ) and number of GCs ( E ) in the spleens of Vav-Cre/ Tet2 +/+ and Vav-Cre/ Tet2 −/− mice. two-tailed t test, ***p

    Journal: Cancer discovery

    Article Title: TET2 deficiency causes germinal center hyperplasia, impairs plasma cell differentiation and promotes B-cell lymphomagenesis

    doi: 10.1158/2159-8290.CD-18-0657

    Figure Lengend Snippet: Tet2 deletion in hematopoietic cells induces GC hyperplasia. A, Representative flow cytometry plot and quantification of (B220 + CD95 + GL7 + ) GC B-cells from Vav-Cre/ Tet2 +/+ (n=5) and Vav-Cre/ Tet2 −/− (n=5) mice at day 10 after immunization with SRBC. B , Quantification of flow cytometry data corresponding to total B cells (B220 + ), mature B cells (B220 + IgD + IgM + ), transitional B cells (B220 + IgD int IgM + ), follicular B cells (B220 + CD23 + CD21 + ), marginal zone B cells (B220 + CD23 low CD21 + ) and plasmablasts/PC (CD138 + ) in the spleens of Vav-Cre/ Tet2 +/+ and Vav-Cre/ Tet2 −/− mice. C , Representative histologic sections of formalin-fixed, paraffin-embedded spleens from Vav-Cre/ Tet2 +/+ and Vav-Cre/ Tet2 −/− mice. Sections were stained with H E and antibodies specific for B220, PNA and Ki-67. D , E , Quantification of GC area ( D ) and number of GCs ( E ) in the spleens of Vav-Cre/ Tet2 +/+ and Vav-Cre/ Tet2 −/− mice. two-tailed t test, ***p

    Article Snippet: Flow cytometry analysis of spleen cell suspensions was performed using the following fluorescent-labeled anti–mouse antibodies: APC-conjugated anti-B220 (BD Biosciences #553092), anti-CD138 (BD Biosciences #558626) and anti-IgM (eBioscience #17–5790-82); PE-conjugated anti-IgD (BD Biosciences #558597), anti-CD23 (eBioscience #5010271) and anti-CD95 (BD Biosciences #554258); PECy7-conjugated anti-CD21 (BioLegend #123420), anti-CD95 (BD Biosciences #557653), anti-GL7 (BD Biosciences # 561530) and anti-CD86 (BD Biosciences #560582); PEVio770-conjugated anti-B220 (Miltenyi Biotec #130–102-308); FITC-conjugated anti-CXCR4 (BD Biosciences #551967) and anti-IgG1 (ebioscience #11–4011-85); Brilliant Violet 421-conjugated anti-CD138 (BioLegend #142507); PerCP-Cy5.5-conjugated anti-GL7 (BioLegend #144610), anti-CD138 (BioLegend #142510) and anti-IgM (BD Biosciences #550881), APC-Cy7-conjugated anti-CD19 (ebioscience #47–0193-82), AlexaFluor647-conjugated anti-BLIMP1 (BD Biosciences #563643).

    Techniques: Flow Cytometry, Mouse Assay, Formalin-fixed Paraffin-Embedded, Staining, Two Tailed Test

    Impaired release of immature bone marrow B cells by CXCR4 antagonism in the absence of S1P1 receptor. (A) Mice were injected with the CXCR4 antagonist AMD3100 or vehicle alone, and bone marrow and blood were collected after 90 min. Immature bone marrow B cells in the blood (B220 + CD93 + ) were gated as IgM + IgD − and IgM + IgD low , and quantified from control and B- S1pr1 KO mice. Results are shown as absolute numbers in 250 µl of blood. Bars represent mean values, and the closed circles are individual mice. Data are representative of three experiments. *, P

    Journal: The Journal of Experimental Medicine

    Article Title: S1P1 receptor directs the release of immature B cells from bone marrow into blood

    doi: 10.1084/jem.20092210

    Figure Lengend Snippet: Impaired release of immature bone marrow B cells by CXCR4 antagonism in the absence of S1P1 receptor. (A) Mice were injected with the CXCR4 antagonist AMD3100 or vehicle alone, and bone marrow and blood were collected after 90 min. Immature bone marrow B cells in the blood (B220 + CD93 + ) were gated as IgM + IgD − and IgM + IgD low , and quantified from control and B- S1pr1 KO mice. Results are shown as absolute numbers in 250 µl of blood. Bars represent mean values, and the closed circles are individual mice. Data are representative of three experiments. *, P

    Article Snippet: Bone marrow cells were labeled with anti-B220 (PE-Cy7 conjugated), anti-IgM (APC conjugated), and anti-IgD (FITC conjugated; BD) and sorted into four populations (B220+ IgM− IgD− , B220+ IgM+ IgD− , B220+ IgM+ IgDlow , and B220+ IgM+ IgDhigh ) using a cell sorter (FACSAria II; BD).

    Techniques: Mouse Assay, Injection

    Migration of bone marrow B cells toward S1P is mediated by the S1P3 receptor. (A) Total bone marrow cells were added to a Transwell insert and allowed to respond to increasing concentrations of S1P or to 100 ng/ml SDF-1 in the lower well. Percentages of the input that were found in the lower well after a 3-h incubation with S1P (left axis) or with SDF-1 (right axis) were plotted for pro–/pre– (B220 + IgD − IgM − ), immature (B220 + IgM + IgD − and B220 + IgM + IgD low ), and mature (B220 + IgM + IgD high ) B cells. Data are presented as mean values ± SD ( n = 3 for each genotype) and are representative of three independent experiments. *, P

    Journal: The Journal of Experimental Medicine

    Article Title: S1P1 receptor directs the release of immature B cells from bone marrow into blood

    doi: 10.1084/jem.20092210

    Figure Lengend Snippet: Migration of bone marrow B cells toward S1P is mediated by the S1P3 receptor. (A) Total bone marrow cells were added to a Transwell insert and allowed to respond to increasing concentrations of S1P or to 100 ng/ml SDF-1 in the lower well. Percentages of the input that were found in the lower well after a 3-h incubation with S1P (left axis) or with SDF-1 (right axis) were plotted for pro–/pre– (B220 + IgD − IgM − ), immature (B220 + IgM + IgD − and B220 + IgM + IgD low ), and mature (B220 + IgM + IgD high ) B cells. Data are presented as mean values ± SD ( n = 3 for each genotype) and are representative of three independent experiments. *, P

    Article Snippet: Bone marrow cells were labeled with anti-B220 (PE-Cy7 conjugated), anti-IgM (APC conjugated), and anti-IgD (FITC conjugated; BD) and sorted into four populations (B220+ IgM− IgD− , B220+ IgM+ IgD− , B220+ IgM+ IgDlow , and B220+ IgM+ IgDhigh ) using a cell sorter (FACSAria II; BD).

    Techniques: Migration, Incubation

    Reduced numbers of mature and immature B cells in the blood of B- S1pr1 KO mice. (A–D) Bone marrow B cell subpopulations from control and B- S1pr1 KO mice were analyzed by flow cytometry using FITC-conjugated anti-B220, APC-conjugated anti-IgM, and PE-conjugated anti-CD43 antibodies. Pro–/pre–B cells were identified as B220 low IgM − , immature B cells were identified as B220 low IgM + , and mature B cells were identified as B220 high IgM + . Pro–B cells were identified as B220 low IgM − CD43 + and pre–B cells were identified as B220 low IgM − CD43 − . Results are shown as density plots (A and C) and as the absolute number of cells counted per femur (B and D). The percentage of cells in each gate is indicated on the plots. Bars represent mean values of pooled data from two experiments, and the closed circles are individual mice. *, P

    Journal: The Journal of Experimental Medicine

    Article Title: S1P1 receptor directs the release of immature B cells from bone marrow into blood

    doi: 10.1084/jem.20092210

    Figure Lengend Snippet: Reduced numbers of mature and immature B cells in the blood of B- S1pr1 KO mice. (A–D) Bone marrow B cell subpopulations from control and B- S1pr1 KO mice were analyzed by flow cytometry using FITC-conjugated anti-B220, APC-conjugated anti-IgM, and PE-conjugated anti-CD43 antibodies. Pro–/pre–B cells were identified as B220 low IgM − , immature B cells were identified as B220 low IgM + , and mature B cells were identified as B220 high IgM + . Pro–B cells were identified as B220 low IgM − CD43 + and pre–B cells were identified as B220 low IgM − CD43 − . Results are shown as density plots (A and C) and as the absolute number of cells counted per femur (B and D). The percentage of cells in each gate is indicated on the plots. Bars represent mean values of pooled data from two experiments, and the closed circles are individual mice. *, P

    Article Snippet: Bone marrow cells were labeled with anti-B220 (PE-Cy7 conjugated), anti-IgM (APC conjugated), and anti-IgD (FITC conjugated; BD) and sorted into four populations (B220+ IgM− IgD− , B220+ IgM+ IgD− , B220+ IgM+ IgDlow , and B220+ IgM+ IgDhigh ) using a cell sorter (FACSAria II; BD).

    Techniques: Mouse Assay, Flow Cytometry, Cytometry

    Immature B cells in the bone marrow do not efficiently enter blood and have elevated apoptosis in the absence of S1P1 receptor. (A–D) Mice were pulsed with BrdU, and B cells from the bone marrow (A and B) and peripheral blood (C and D) of control and B- S1pr1 KO mice were analyzed by flow cytometry using anti-B220, anti-IgD, and anti-IgM antibodies in combination with BrdU detection methodology, as described in Materials and methods. Results are shown as dot plots (A and C), and as absolute numbers of BrdU + B220 low IgM − (pro–/pre–) and B220 low IgM + (immature) B cells per femur (B) and BrdU + B220 + IgD low IgM high (immature) and B220 + IgD high IgM low (mature) B cells per 400 µl of blood (D). The percentage of cells in each gate is indicated on the plots. Bars represent mean values, and the closed circles are individual mice. Data are representative of three experiments with three to five mice of each genotype per experiment. *, P

    Journal: The Journal of Experimental Medicine

    Article Title: S1P1 receptor directs the release of immature B cells from bone marrow into blood

    doi: 10.1084/jem.20092210

    Figure Lengend Snippet: Immature B cells in the bone marrow do not efficiently enter blood and have elevated apoptosis in the absence of S1P1 receptor. (A–D) Mice were pulsed with BrdU, and B cells from the bone marrow (A and B) and peripheral blood (C and D) of control and B- S1pr1 KO mice were analyzed by flow cytometry using anti-B220, anti-IgD, and anti-IgM antibodies in combination with BrdU detection methodology, as described in Materials and methods. Results are shown as dot plots (A and C), and as absolute numbers of BrdU + B220 low IgM − (pro–/pre–) and B220 low IgM + (immature) B cells per femur (B) and BrdU + B220 + IgD low IgM high (immature) and B220 + IgD high IgM low (mature) B cells per 400 µl of blood (D). The percentage of cells in each gate is indicated on the plots. Bars represent mean values, and the closed circles are individual mice. Data are representative of three experiments with three to five mice of each genotype per experiment. *, P

    Article Snippet: Bone marrow cells were labeled with anti-B220 (PE-Cy7 conjugated), anti-IgM (APC conjugated), and anti-IgD (FITC conjugated; BD) and sorted into four populations (B220+ IgM− IgD− , B220+ IgM+ IgD− , B220+ IgM+ IgDlow , and B220+ IgM+ IgDhigh ) using a cell sorter (FACSAria II; BD).

    Techniques: Mouse Assay, Flow Cytometry, Cytometry

    CD69 expression in bone marrow B cells modulates the appearance of immature B cells in peripheral blood. (A) CD69 expression on bone marrow B cells. Results are shown as anti-CD69 fluorescence intensity for cells from control and B- S1pr1 KO mice compared with the isotype control. Representative results are from nine independent experiments. (B) Expression of the human CD69 transgene on bone marrow total B cells in control and transgenic (CD69-Tg) mice. B220 + bone marrow cells were analyzed for their expression of human CD69 by flow cytometry. (C and D) Distribution of B220 low IgM − (pro–/pre–) and B220 low IgM + (immature) B cells in the bone marrow (C) and of B220 + IgD low IgM + (immature) and B220 + IgD high IgM low (mature) B cells in the peripheral blood (D) of control and CD69 transgenic mice. Bars represent mean values, and the closed circles are individual mice. Data represent pooled results from three experiments. **, P

    Journal: The Journal of Experimental Medicine

    Article Title: S1P1 receptor directs the release of immature B cells from bone marrow into blood

    doi: 10.1084/jem.20092210

    Figure Lengend Snippet: CD69 expression in bone marrow B cells modulates the appearance of immature B cells in peripheral blood. (A) CD69 expression on bone marrow B cells. Results are shown as anti-CD69 fluorescence intensity for cells from control and B- S1pr1 KO mice compared with the isotype control. Representative results are from nine independent experiments. (B) Expression of the human CD69 transgene on bone marrow total B cells in control and transgenic (CD69-Tg) mice. B220 + bone marrow cells were analyzed for their expression of human CD69 by flow cytometry. (C and D) Distribution of B220 low IgM − (pro–/pre–) and B220 low IgM + (immature) B cells in the bone marrow (C) and of B220 + IgD low IgM + (immature) and B220 + IgD high IgM low (mature) B cells in the peripheral blood (D) of control and CD69 transgenic mice. Bars represent mean values, and the closed circles are individual mice. Data represent pooled results from three experiments. **, P

    Article Snippet: Bone marrow cells were labeled with anti-B220 (PE-Cy7 conjugated), anti-IgM (APC conjugated), and anti-IgD (FITC conjugated; BD) and sorted into four populations (B220+ IgM− IgD− , B220+ IgM+ IgD− , B220+ IgM+ IgDlow , and B220+ IgM+ IgDhigh ) using a cell sorter (FACSAria II; BD).

    Techniques: Expressing, Fluorescence, Mouse Assay, Transgenic Assay, Flow Cytometry, Cytometry

    SVP[Rapa] treatment enables AAV8 re-administration in nonhuman primates. a Protocol outline. Male naive cynomolgus monkeys were treated i.v. with 2 × 10 12 vg kg −1 of AAV8-Gaa vector and either SVP[Rapa] (3 mg kg −1 , n = 2, SVP[Rapa]#1 and SVP[Rapa]#2) or SVP[empty] control ( n = 1) and then challenged i.v. on day 30 with 2 × 10 12 vg kg − 1 of AAV8-hF.IX vector and either SVP[Rapa] or SVP[empty] control, as described above. b , c Analysis of b anti-AAV8 IgG antibodies and c anti-AAV8 IgM antibody responses measured by ELISA. d Analysis of anti-AAV8 neutralizing antibodies (NAb) measured with a cell-based neutralization assay. e Analysis of anti-AAV8 IgG secreting B cells in splenocytes measured by B ELISpot. Data are shown as individual replicates and the bars represent mean ± s.d. The dotted line indicates the threshold for positivity corresponding to 50 spot forming units (SFU) per million cells. f Plasma hF.IX antigen levels quantified by ELISA at the indicated time points following administration of AAV8-hF.IX vector. g AAV8-hF.IX vector genome copy number (VGCN) per diploid genome in liver. The symbols represent individual liver lobes (left, right, caudate and quadrate) and the bars represent the mean ± s.d. (4 liver lobes per monkey; one-way ANOVA with Tukey post hoc test, ** p

    Journal: Nature Communications

    Article Title: Antigen-selective modulation of AAV immunogenicity with tolerogenic rapamycin nanoparticles enables successful vector re-administration

    doi: 10.1038/s41467-018-06621-3

    Figure Lengend Snippet: SVP[Rapa] treatment enables AAV8 re-administration in nonhuman primates. a Protocol outline. Male naive cynomolgus monkeys were treated i.v. with 2 × 10 12 vg kg −1 of AAV8-Gaa vector and either SVP[Rapa] (3 mg kg −1 , n = 2, SVP[Rapa]#1 and SVP[Rapa]#2) or SVP[empty] control ( n = 1) and then challenged i.v. on day 30 with 2 × 10 12 vg kg − 1 of AAV8-hF.IX vector and either SVP[Rapa] or SVP[empty] control, as described above. b , c Analysis of b anti-AAV8 IgG antibodies and c anti-AAV8 IgM antibody responses measured by ELISA. d Analysis of anti-AAV8 neutralizing antibodies (NAb) measured with a cell-based neutralization assay. e Analysis of anti-AAV8 IgG secreting B cells in splenocytes measured by B ELISpot. Data are shown as individual replicates and the bars represent mean ± s.d. The dotted line indicates the threshold for positivity corresponding to 50 spot forming units (SFU) per million cells. f Plasma hF.IX antigen levels quantified by ELISA at the indicated time points following administration of AAV8-hF.IX vector. g AAV8-hF.IX vector genome copy number (VGCN) per diploid genome in liver. The symbols represent individual liver lobes (left, right, caudate and quadrate) and the bars represent the mean ± s.d. (4 liver lobes per monkey; one-way ANOVA with Tukey post hoc test, ** p

    Article Snippet: For NHP samples, anti-monkey IgG-HRP (dilution 1:20,000, 43R-IG020HRP, Fitzgerald Industries, Acton, MA) and anti-IgM (dilution 1:5000, 2020-05, Southern Biotech) were used.

    Techniques: Plasmid Preparation, Enzyme-linked Immunosorbent Assay, Neutralization, Enzyme-linked Immunospot

    Co-expression of sortilin, APLP2, or both with PCSK9 has no major effect on LDLR degradation. Huh7 cells were transfected with a total of 3 μg using 1 μg of each vector encoding for either a control protein 7B2 (−), sortilin (+), APLP2 (+), or PCSK9 (+), as indicated. After 48 h, lysates were analyzed by Western blotting for expression of the LDLR, sortilin-Myc, APLP2-V5, intracellular pro- and mature-PCSK9-V5, and β-actin. Media were analyzed for secreted endogenous and overexpressed PCSK9-V5 using a rabbit polyclonal human PCSK9 antibody. Quantification of LDLR expression was normalized against that of β-actin. These data are representative of at least 3 different experiments showing similar results.

    Journal: The Journal of Biological Chemistry

    Article Title: Amyloid Precursor-like Protein 2 and Sortilin Do Not Regulate the PCSK9 Convertase-mediated Low Density Lipoprotein Receptor Degradation but Interact with Each Other *

    doi: 10.1074/jbc.M115.647180

    Figure Lengend Snippet: Co-expression of sortilin, APLP2, or both with PCSK9 has no major effect on LDLR degradation. Huh7 cells were transfected with a total of 3 μg using 1 μg of each vector encoding for either a control protein 7B2 (−), sortilin (+), APLP2 (+), or PCSK9 (+), as indicated. After 48 h, lysates were analyzed by Western blotting for expression of the LDLR, sortilin-Myc, APLP2-V5, intracellular pro- and mature-PCSK9-V5, and β-actin. Media were analyzed for secreted endogenous and overexpressed PCSK9-V5 using a rabbit polyclonal human PCSK9 antibody. Quantification of LDLR expression was normalized against that of β-actin. These data are representative of at least 3 different experiments showing similar results.

    Article Snippet: Mouse APLP2 was detected using a rabbit polyclonal antibody kindly provided by Dr. G. Thinakaran (University of Chicago), whereas human APLP2 was detected with either mAb-V5 or a rabbit polyclonal antibody (Novus Biologicals).

    Techniques: Expressing, Transfection, Plasmid Preparation, Western Blot

    Co-localization of PAB-reactive PLTA antigen, IgA, IgM, and C3c detected by double fluorescence immunohistochemistry. A: Anti-IgA antibody (red) vs PAB antibody (green) after MT treatment, B: anti-IgM antibody (red) vs PAB antibody (green) after MT treatment, C: anti-IgM antibody (red) vs anti-IgA antibody (green), and D: anti-C3c antibody (red) vs PAB antibody (green) after MT treatment. Many PAB-reactive SRBs were also positive for IgA, IgM, and C3c, showing yellow-colored double-positive signals (A, B, and D, respectively). Both IgA and IgM colocalized with these SRBs, indicated by yellow-colored double-positive signals (C).

    Journal: PLoS ONE

    Article Title: Propionibacterium acnes-derived insoluble immune complexes in sinus macrophages of lymph nodes affected by sarcoidosis

    doi: 10.1371/journal.pone.0192408

    Figure Lengend Snippet: Co-localization of PAB-reactive PLTA antigen, IgA, IgM, and C3c detected by double fluorescence immunohistochemistry. A: Anti-IgA antibody (red) vs PAB antibody (green) after MT treatment, B: anti-IgM antibody (red) vs PAB antibody (green) after MT treatment, C: anti-IgM antibody (red) vs anti-IgA antibody (green), and D: anti-C3c antibody (red) vs PAB antibody (green) after MT treatment. Many PAB-reactive SRBs were also positive for IgA, IgM, and C3c, showing yellow-colored double-positive signals (A, B, and D, respectively). Both IgA and IgM colocalized with these SRBs, indicated by yellow-colored double-positive signals (C).

    Article Snippet: After the initial incubation, the plates were further incubated for 60 min at 37°C with unlabeled mouse anti-PLTA antibody (PAB) diluted 1:500; unlabeled rabbit anti-human C1q and C3c (DAKO 0136 and A062, respectively) diluted 1:500; or biotinylated goat anti-human IgG, IgA, or IgM antibody (Invitrogen, Carlsbad, CA, USA) diluted 1:5000.

    Techniques: Fluorescence, Immunohistochemistry

    Immuno-electron microscopy images of SRBs in sinus macrophages of sarcoid lymph nodes. A and B: IHC with PAB antibody after MT treatment, C: IHC with anti-IgA antibody, and D: IHC with anti-IgM antibody. Note that dense black-colored reaction products by each antibody were located along the peripheral rim of the SRBs. A similar distribution of PAB-reactivity was observed in a large spherical-shaped HW body (A).

    Journal: PLoS ONE

    Article Title: Propionibacterium acnes-derived insoluble immune complexes in sinus macrophages of lymph nodes affected by sarcoidosis

    doi: 10.1371/journal.pone.0192408

    Figure Lengend Snippet: Immuno-electron microscopy images of SRBs in sinus macrophages of sarcoid lymph nodes. A and B: IHC with PAB antibody after MT treatment, C: IHC with anti-IgA antibody, and D: IHC with anti-IgM antibody. Note that dense black-colored reaction products by each antibody were located along the peripheral rim of the SRBs. A similar distribution of PAB-reactivity was observed in a large spherical-shaped HW body (A).

    Article Snippet: After the initial incubation, the plates were further incubated for 60 min at 37°C with unlabeled mouse anti-PLTA antibody (PAB) diluted 1:500; unlabeled rabbit anti-human C1q and C3c (DAKO 0136 and A062, respectively) diluted 1:500; or biotinylated goat anti-human IgG, IgA, or IgM antibody (Invitrogen, Carlsbad, CA, USA) diluted 1:5000.

    Techniques: Immuno-Electron Microscopy, Immunohistochemistry

    Insoluble immune complexes in sinus macrophages of control lymph nodes from colon cancer patients. In a representative case of control lymph nodes with IICs from colon cancer patients, identical areas of the lymphatic sinus are shown in semi-serial sections; HE stain (A), IHC with anti-human IgG (B), IgA (C), and IgM (D) antibody, IHC with PAB antibody (E), and IHC with PAB antibody after MT treatment (F). In the sinus macrophages, many IgA-positive and a few IgM-positive small particles were detected (C and D, respectively), although a few PAB-reactive SRBs were detected with no difference in the number between the sections with and without MT treatment (F and E, respectively). Scale bar: 20 μm.

    Journal: PLoS ONE

    Article Title: Propionibacterium acnes-derived insoluble immune complexes in sinus macrophages of lymph nodes affected by sarcoidosis

    doi: 10.1371/journal.pone.0192408

    Figure Lengend Snippet: Insoluble immune complexes in sinus macrophages of control lymph nodes from colon cancer patients. In a representative case of control lymph nodes with IICs from colon cancer patients, identical areas of the lymphatic sinus are shown in semi-serial sections; HE stain (A), IHC with anti-human IgG (B), IgA (C), and IgM (D) antibody, IHC with PAB antibody (E), and IHC with PAB antibody after MT treatment (F). In the sinus macrophages, many IgA-positive and a few IgM-positive small particles were detected (C and D, respectively), although a few PAB-reactive SRBs were detected with no difference in the number between the sections with and without MT treatment (F and E, respectively). Scale bar: 20 μm.

    Article Snippet: After the initial incubation, the plates were further incubated for 60 min at 37°C with unlabeled mouse anti-PLTA antibody (PAB) diluted 1:500; unlabeled rabbit anti-human C1q and C3c (DAKO 0136 and A062, respectively) diluted 1:500; or biotinylated goat anti-human IgG, IgA, or IgM antibody (Invitrogen, Carlsbad, CA, USA) diluted 1:5000.

    Techniques: H&E Stain, Immunohistochemistry

    Insoluble immune complexes in sinus macrophages of control lymph nodes from patients with reactive lymphadenitis. In a representative case of control lymph nodes with IICs from patients with reactive lymphadenitis, identical areas of the lymphatic sinus are shown in semi-serial sections; HE stain (A), IHC with anti-human IgG (B), IgA (C), and IgM (D) antibody, IHC with PAB antibody (E), and IHC with PAB antibody after MT treatment (F). In the sinus macrophages, IgA- and IgM-positive SRBs were detected (C and D, respectively), and PAB-reactive SRBs were also detected with a small increase in the number after MT treatment (F). Scale bar: 20 μm.

    Journal: PLoS ONE

    Article Title: Propionibacterium acnes-derived insoluble immune complexes in sinus macrophages of lymph nodes affected by sarcoidosis

    doi: 10.1371/journal.pone.0192408

    Figure Lengend Snippet: Insoluble immune complexes in sinus macrophages of control lymph nodes from patients with reactive lymphadenitis. In a representative case of control lymph nodes with IICs from patients with reactive lymphadenitis, identical areas of the lymphatic sinus are shown in semi-serial sections; HE stain (A), IHC with anti-human IgG (B), IgA (C), and IgM (D) antibody, IHC with PAB antibody (E), and IHC with PAB antibody after MT treatment (F). In the sinus macrophages, IgA- and IgM-positive SRBs were detected (C and D, respectively), and PAB-reactive SRBs were also detected with a small increase in the number after MT treatment (F). Scale bar: 20 μm.

    Article Snippet: After the initial incubation, the plates were further incubated for 60 min at 37°C with unlabeled mouse anti-PLTA antibody (PAB) diluted 1:500; unlabeled rabbit anti-human C1q and C3c (DAKO 0136 and A062, respectively) diluted 1:500; or biotinylated goat anti-human IgG, IgA, or IgM antibody (Invitrogen, Carlsbad, CA, USA) diluted 1:5000.

    Techniques: H&E Stain, Immunohistochemistry

    P . acnes -derived insoluble immune complexes in sinus macrophages of sarcoid lymph nodes. In a representative case of sarcoid lymph nodes, identical areas of the lesion including a lymphatic sinus and adjacent paracortical area with a sarcoid granuloma are shown in semi-serial sections; HE stain (A), IHC with anti-human IgG (B), IgA (C), and IgM (D) antibody, IHC with PAB antibody (E), and IHC with PAB antibody after MT treatment (F). In the sinus macrophages, the distributions of IgA- and IgM-positive SRBs (C and D) and PAB-reactive SRBs (F) were similar. Note the few PAB-reactive SRBs with weak intensity (indicated by the arrow) in the granuloma after MT treatment (F). Scale bar: 20 μm.

    Journal: PLoS ONE

    Article Title: Propionibacterium acnes-derived insoluble immune complexes in sinus macrophages of lymph nodes affected by sarcoidosis

    doi: 10.1371/journal.pone.0192408

    Figure Lengend Snippet: P . acnes -derived insoluble immune complexes in sinus macrophages of sarcoid lymph nodes. In a representative case of sarcoid lymph nodes, identical areas of the lesion including a lymphatic sinus and adjacent paracortical area with a sarcoid granuloma are shown in semi-serial sections; HE stain (A), IHC with anti-human IgG (B), IgA (C), and IgM (D) antibody, IHC with PAB antibody (E), and IHC with PAB antibody after MT treatment (F). In the sinus macrophages, the distributions of IgA- and IgM-positive SRBs (C and D) and PAB-reactive SRBs (F) were similar. Note the few PAB-reactive SRBs with weak intensity (indicated by the arrow) in the granuloma after MT treatment (F). Scale bar: 20 μm.

    Article Snippet: After the initial incubation, the plates were further incubated for 60 min at 37°C with unlabeled mouse anti-PLTA antibody (PAB) diluted 1:500; unlabeled rabbit anti-human C1q and C3c (DAKO 0136 and A062, respectively) diluted 1:500; or biotinylated goat anti-human IgG, IgA, or IgM antibody (Invitrogen, Carlsbad, CA, USA) diluted 1:5000.

    Techniques: Derivative Assay, H&E Stain, Immunohistochemistry