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  • 99
    Vector Laboratories biotinylated secondary antibodies
    Biotinylated Secondary Antibodies, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 99/100, based on 10949 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher goat anti rabbit igg h l cross adsorbed secondary antibody
    Goat Anti Rabbit Igg H L Cross Adsorbed Secondary Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 14192 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher goat anti mouse igg h l cross adsorbed secondary antibody
    Goat Anti Mouse Igg H L Cross Adsorbed Secondary Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 14005 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher goat anti rabbit igg h l highly cross adsorbed secondary antibody
    Goat Anti Rabbit Igg H L Highly Cross Adsorbed Secondary Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 11348 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher donkey anti rabbit igg h l highly cross adsorbed secondary antibody
    Donkey Anti Rabbit Igg H L Highly Cross Adsorbed Secondary Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 9653 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher goat anti mouse igg h l highly cross adsorbed secondary antibody
    Goat Anti Mouse Igg H L Highly Cross Adsorbed Secondary Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 9004 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore igg
    Igg, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 10101 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher igg
    Igg, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 9957 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher donkey anti mouse igg h l highly cross adsorbed secondary antibody
    Donkey Anti Mouse Igg H L Highly Cross Adsorbed Secondary Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 6700 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    SouthernBiotech igg4
    Antigen-specific <t>IgG</t> subclasses during follow-up and relapse. The PLA 2 R1- (A) and THSD7A-specific (B) IgG subclasses did not significantly change during the follow-up period. At the time of relapse of PLA 2 R1-ab, antigen-specific IgG subclasses were similar to the distribution observed at baseline (C) .
    Igg4, supplied by SouthernBiotech, used in various techniques. Bioz Stars score: 95/100, based on 279 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Becton Dickinson igg3
    GaMD immune serum from WT mice inhibits GaMD-induced renal disease without decreasing other isotypes if injected into GaMD-immunized γ1 - mice by 5d after immunization a . BALB/c γ1 - mice (4 or 8/gp) were injected s.c. with GaMD on day 0 and i.p. with 0.5 ml of pooled serum from GaMD-immunized WT mice (GaMD immune WT serum) or unimmunized WT mice (non-immune serum), starting 4, 5, or 6d after GaMD immunization. Day 7 urine samples were analyzed. ND = none detected. b. BALB/c γ1 - mice (4 or 8/gp) were injected s.c. with GaMD on day 0 and i.p. with 0.5 ml of pooled serum from GaMD-immunized WT mice (GaMD immune WT serum) or unimmunized WT mice (non-immune serum), 5, 6 and 7 d after GaMD immunization. Sera were assayed for total IgG1, IgG2a, IgM and <t>IgG3</t> on d0 (unimmunized) and 8d after GaMD immunization. ND = none detected.For both a and b , * indicates p
    Igg3, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 94/100, based on 865 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    98
    Cell Signaling Technology Inc anti rabbit igg
    GaMD immune serum from WT mice inhibits GaMD-induced renal disease without decreasing other isotypes if injected into GaMD-immunized γ1 - mice by 5d after immunization a . BALB/c γ1 - mice (4 or 8/gp) were injected s.c. with GaMD on day 0 and i.p. with 0.5 ml of pooled serum from GaMD-immunized WT mice (GaMD immune WT serum) or unimmunized WT mice (non-immune serum), starting 4, 5, or 6d after GaMD immunization. Day 7 urine samples were analyzed. ND = none detected. b. BALB/c γ1 - mice (4 or 8/gp) were injected s.c. with GaMD on day 0 and i.p. with 0.5 ml of pooled serum from GaMD-immunized WT mice (GaMD immune WT serum) or unimmunized WT mice (non-immune serum), 5, 6 and 7 d after GaMD immunization. Sera were assayed for total IgG1, IgG2a, IgM and <t>IgG3</t> on d0 (unimmunized) and 8d after GaMD immunization. ND = none detected.For both a and b , * indicates p
    Anti Rabbit Igg, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 98/100, based on 6668 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Vector Laboratories biotinylated goat anti rabbit igg
    GaMD immune serum from WT mice inhibits GaMD-induced renal disease without decreasing other isotypes if injected into GaMD-immunized γ1 - mice by 5d after immunization a . BALB/c γ1 - mice (4 or 8/gp) were injected s.c. with GaMD on day 0 and i.p. with 0.5 ml of pooled serum from GaMD-immunized WT mice (GaMD immune WT serum) or unimmunized WT mice (non-immune serum), starting 4, 5, or 6d after GaMD immunization. Day 7 urine samples were analyzed. ND = none detected. b. BALB/c γ1 - mice (4 or 8/gp) were injected s.c. with GaMD on day 0 and i.p. with 0.5 ml of pooled serum from GaMD-immunized WT mice (GaMD immune WT serum) or unimmunized WT mice (non-immune serum), 5, 6 and 7 d after GaMD immunization. Sera were assayed for total IgG1, IgG2a, IgM and <t>IgG3</t> on d0 (unimmunized) and 8d after GaMD immunization. ND = none detected.For both a and b , * indicates p
    Biotinylated Goat Anti Rabbit Igg, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 99/100, based on 9049 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam goat anti rabbit igg h l hrp
    GaMD immune serum from WT mice inhibits GaMD-induced renal disease without decreasing other isotypes if injected into GaMD-immunized γ1 - mice by 5d after immunization a . BALB/c γ1 - mice (4 or 8/gp) were injected s.c. with GaMD on day 0 and i.p. with 0.5 ml of pooled serum from GaMD-immunized WT mice (GaMD immune WT serum) or unimmunized WT mice (non-immune serum), starting 4, 5, or 6d after GaMD immunization. Day 7 urine samples were analyzed. ND = none detected. b. BALB/c γ1 - mice (4 or 8/gp) were injected s.c. with GaMD on day 0 and i.p. with 0.5 ml of pooled serum from GaMD-immunized WT mice (GaMD immune WT serum) or unimmunized WT mice (non-immune serum), 5, 6 and 7 d after GaMD immunization. Sera were assayed for total IgG1, IgG2a, IgM and <t>IgG3</t> on d0 (unimmunized) and 8d after GaMD immunization. ND = none detected.For both a and b , * indicates p
    Goat Anti Rabbit Igg H L Hrp, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 2991 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore rabbit igg
    GaMD immune serum from WT mice inhibits GaMD-induced renal disease without decreasing other isotypes if injected into GaMD-immunized γ1 - mice by 5d after immunization a . BALB/c γ1 - mice (4 or 8/gp) were injected s.c. with GaMD on day 0 and i.p. with 0.5 ml of pooled serum from GaMD-immunized WT mice (GaMD immune WT serum) or unimmunized WT mice (non-immune serum), starting 4, 5, or 6d after GaMD immunization. Day 7 urine samples were analyzed. ND = none detected. b. BALB/c γ1 - mice (4 or 8/gp) were injected s.c. with GaMD on day 0 and i.p. with 0.5 ml of pooled serum from GaMD-immunized WT mice (GaMD immune WT serum) or unimmunized WT mice (non-immune serum), 5, 6 and 7 d after GaMD immunization. Sera were assayed for total IgG1, IgG2a, IgM and <t>IgG3</t> on d0 (unimmunized) and 8d after GaMD immunization. ND = none detected.For both a and b , * indicates p
    Rabbit Igg, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 7847 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher mouse igg1
    Lack of C1q affects the level of immunoglobulins IgA (A) , <t>IgG1</t> (B) , IgG2a (C) , IgG2b (D) , IgG3 (E) , and IgM (F) in infected mice. Whole blood was collected from mice infected with 10 4 Borrelia burgdorferi ) at 7, 10, 21, and 28 days post-infection. Serum level of immunoglobulins was determined by the Bio-Plex system employing the Luminex multianalyte profiling technology. White circles refer to samples from C57BL/6 mice, whereas red squares represent data from C1qα −/− mice ( n = 5; * P
    Mouse Igg1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 1175 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Santa Cruz Biotechnology normal rabbit igg
    GABA degrading enzyme is expressed in rat kidney. A) RT-PCR results for ABAT in rat kidney cortex. Appropriately-sized bands were detected for ABAT in at least five independent experiments. Rat brain RNA or commercial universal RNA was used as positive control. M: molecular marker. B) Immunoblotting for ABAT in rat kidney cortex. B: brain control, K: kidney cortex. C) Immunostaining for ABAT in rat kidney cortex. Staining with normal <t>IgG</t> instead of primary antibody served as negative control.
    Normal Rabbit Igg, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 94/100, based on 7871 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore anti rabbit igg whole molecule peroxidase antibody
    GABA degrading enzyme is expressed in rat kidney. A) RT-PCR results for ABAT in rat kidney cortex. Appropriately-sized bands were detected for ABAT in at least five independent experiments. Rat brain RNA or commercial universal RNA was used as positive control. M: molecular marker. B) Immunoblotting for ABAT in rat kidney cortex. B: brain control, K: kidney cortex. C) Immunostaining for ABAT in rat kidney cortex. Staining with normal <t>IgG</t> instead of primary antibody served as negative control.
    Anti Rabbit Igg Whole Molecule Peroxidase Antibody, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 7032 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher anti mouse igg
    GABA degrading enzyme is expressed in rat kidney. A) RT-PCR results for ABAT in rat kidney cortex. Appropriately-sized bands were detected for ABAT in at least five independent experiments. Rat brain RNA or commercial universal RNA was used as positive control. M: molecular marker. B) Immunoblotting for ABAT in rat kidney cortex. B: brain control, K: kidney cortex. C) Immunostaining for ABAT in rat kidney cortex. Staining with normal <t>IgG</t> instead of primary antibody served as negative control.
    Anti Mouse Igg, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 6123 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore anti mouse igg whole molecule peroxidase antibody
    GABA degrading enzyme is expressed in rat kidney. A) RT-PCR results for ABAT in rat kidney cortex. Appropriately-sized bands were detected for ABAT in at least five independent experiments. Rat brain RNA or commercial universal RNA was used as positive control. M: molecular marker. B) Immunoblotting for ABAT in rat kidney cortex. B: brain control, K: kidney cortex. C) Immunostaining for ABAT in rat kidney cortex. Staining with normal <t>IgG</t> instead of primary antibody served as negative control.
    Anti Mouse Igg Whole Molecule Peroxidase Antibody, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 5324 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    N/A
    Pig polyclonal to IgG1 IgG2 IgG3 IgG4 Isotype Note IgG Host Note Pig Conjugation Note Unconjugated Reactivity Note Human
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    Sheep Serum anti Human IgG1 IgG2 IgG3 IgG4 subclass specific Host Species Note Sheep Human
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    Image Search Results


    Antigen-specific IgG subclasses during follow-up and relapse. The PLA 2 R1- (A) and THSD7A-specific (B) IgG subclasses did not significantly change during the follow-up period. At the time of relapse of PLA 2 R1-ab, antigen-specific IgG subclasses were similar to the distribution observed at baseline (C) .

    Journal: Frontiers in Immunology

    Article Title: Antigen-Specific IgG Subclasses in Primary and Malignancy-Associated Membranous Nephropathy

    doi: 10.3389/fimmu.2018.03035

    Figure Lengend Snippet: Antigen-specific IgG subclasses during follow-up and relapse. The PLA 2 R1- (A) and THSD7A-specific (B) IgG subclasses did not significantly change during the follow-up period. At the time of relapse of PLA 2 R1-ab, antigen-specific IgG subclasses were similar to the distribution observed at baseline (C) .

    Article Snippet: For detection of subclass-specific IgG in Western blot analyses, horseradish peroxidase-conjugated monoclonal anti-human IgG1 (9054-05), IgG2 (9060-05), IgG3 (9210-05), IgG4 (9200-05), and IgG (9040-05) antibodies from mouse were used (IgG1, IgG2, and IgG3 at a 1:5,000 dilution; IgG4 at a 1:10,000 dilution; IgG at a 1:20,000 dilution, all SouthernBiotech, Birmingham, USA), diluted in 5% dry milk in PBS-T.

    Techniques: Proximity Ligation Assay

    Representative Western blots of patients with primary MN and malignancy-associated MN. Representative Western blot analyses of patients with all 4 subclasses of antigen-specific IgG binding. (A) Patient 1 with PLA 2 R1-associated MN and malignancy. (B) Patient 2 with PLA 2 R1-associated MN and no malignancy. (C) Patient 3 with THSD7A-associated MN and malignancy. (D) Patient 4 with THSD7A-associated MN and no malignancy.

    Journal: Frontiers in Immunology

    Article Title: Antigen-Specific IgG Subclasses in Primary and Malignancy-Associated Membranous Nephropathy

    doi: 10.3389/fimmu.2018.03035

    Figure Lengend Snippet: Representative Western blots of patients with primary MN and malignancy-associated MN. Representative Western blot analyses of patients with all 4 subclasses of antigen-specific IgG binding. (A) Patient 1 with PLA 2 R1-associated MN and malignancy. (B) Patient 2 with PLA 2 R1-associated MN and no malignancy. (C) Patient 3 with THSD7A-associated MN and malignancy. (D) Patient 4 with THSD7A-associated MN and no malignancy.

    Article Snippet: For detection of subclass-specific IgG in Western blot analyses, horseradish peroxidase-conjugated monoclonal anti-human IgG1 (9054-05), IgG2 (9060-05), IgG3 (9210-05), IgG4 (9200-05), and IgG (9040-05) antibodies from mouse were used (IgG1, IgG2, and IgG3 at a 1:5,000 dilution; IgG4 at a 1:10,000 dilution; IgG at a 1:20,000 dilution, all SouthernBiotech, Birmingham, USA), diluted in 5% dry milk in PBS-T.

    Techniques: Western Blot, Binding Assay, Proximity Ligation Assay

    Binding of antigen-specific IgG subclasses to human glomerular and lung tissue. Representative Western blot analyses of the antigen-specific IgG subclasses binding to lung tissue extracts (LTE) and human glomerular extracts (HGE). (A) Serum from patient 1 was positive for all THSD7A-specific IgG subclasses. (B) Serum from patient 2 showed detectability for THSD7A-specific IgG3 and IgG4. (C) Serum from patient 3 was positive for PLA 2 R1-specific IgG3 and IgG4.

    Journal: Frontiers in Immunology

    Article Title: Antigen-Specific IgG Subclasses in Primary and Malignancy-Associated Membranous Nephropathy

    doi: 10.3389/fimmu.2018.03035

    Figure Lengend Snippet: Binding of antigen-specific IgG subclasses to human glomerular and lung tissue. Representative Western blot analyses of the antigen-specific IgG subclasses binding to lung tissue extracts (LTE) and human glomerular extracts (HGE). (A) Serum from patient 1 was positive for all THSD7A-specific IgG subclasses. (B) Serum from patient 2 showed detectability for THSD7A-specific IgG3 and IgG4. (C) Serum from patient 3 was positive for PLA 2 R1-specific IgG3 and IgG4.

    Article Snippet: For detection of subclass-specific IgG in Western blot analyses, horseradish peroxidase-conjugated monoclonal anti-human IgG1 (9054-05), IgG2 (9060-05), IgG3 (9210-05), IgG4 (9200-05), and IgG (9040-05) antibodies from mouse were used (IgG1, IgG2, and IgG3 at a 1:5,000 dilution; IgG4 at a 1:10,000 dilution; IgG at a 1:20,000 dilution, all SouthernBiotech, Birmingham, USA), diluted in 5% dry milk in PBS-T.

    Techniques: Binding Assay, Western Blot, Proximity Ligation Assay

    THSD7A and PLA 2 R1 are differentially glycosylated in human lung and kidney. (A) The THSD7A-ab positive serum shows positive binding to the antigen of human lung tissue (LTE) and glomerular extract (HGE), however, the antigen migrates differentially in both tissues. The renal antigen showed a higher molecular weight compared to the lung antigen. (B) Upon addition of deglycosylating enzymes THSD7A shows the same migration pattern in both tissues, suggesting a polypeptide backbone with different glycosylation in the lung and the kidney. (C,D) Similar observations were made for PLA 2 R1 in lung and kidney tissue. 140 kDa band in LTE represents residual IgG, which was also detected in negative controls (healthy serum, n = 5, data not shown).

    Journal: Frontiers in Immunology

    Article Title: Antigen-Specific IgG Subclasses in Primary and Malignancy-Associated Membranous Nephropathy

    doi: 10.3389/fimmu.2018.03035

    Figure Lengend Snippet: THSD7A and PLA 2 R1 are differentially glycosylated in human lung and kidney. (A) The THSD7A-ab positive serum shows positive binding to the antigen of human lung tissue (LTE) and glomerular extract (HGE), however, the antigen migrates differentially in both tissues. The renal antigen showed a higher molecular weight compared to the lung antigen. (B) Upon addition of deglycosylating enzymes THSD7A shows the same migration pattern in both tissues, suggesting a polypeptide backbone with different glycosylation in the lung and the kidney. (C,D) Similar observations were made for PLA 2 R1 in lung and kidney tissue. 140 kDa band in LTE represents residual IgG, which was also detected in negative controls (healthy serum, n = 5, data not shown).

    Article Snippet: For detection of subclass-specific IgG in Western blot analyses, horseradish peroxidase-conjugated monoclonal anti-human IgG1 (9054-05), IgG2 (9060-05), IgG3 (9210-05), IgG4 (9200-05), and IgG (9040-05) antibodies from mouse were used (IgG1, IgG2, and IgG3 at a 1:5,000 dilution; IgG4 at a 1:10,000 dilution; IgG at a 1:20,000 dilution, all SouthernBiotech, Birmingham, USA), diluted in 5% dry milk in PBS-T.

    Techniques: Proximity Ligation Assay, Binding Assay, Molecular Weight, Migration

    Distribution of PLA 2 R1- and THSD7A-specific IgG subclasses at baseline. (A) Detection of antigen-specific IgG subclasses in the complete study cohort by Western Blot using recombinant proteins. All patients were found to be positive for antigen-specific IgG4-antibodies. Antigen-specific IgG3 antibodies were more common than antigen-specific IgG1, while IgG2 was detected less often. The distribution of (B) PLA 2 R1- and (C) THSD7A-specific IgG subclasses was not different in patients with or without malignancy. Patients with (D) PLA 2 R1- or (E) THSD7A-associated MN were grouped depending on the clinical outcome: negativity of autoantibodies during follow-up, remission of proteinuria, doubling of serum creatinine. IgG1-3 depicts the percentage of patients in whom at least one antigen-specific IgG subclass (IgG1, IgG2, or IgG3) was detected in addition to IgG4.

    Journal: Frontiers in Immunology

    Article Title: Antigen-Specific IgG Subclasses in Primary and Malignancy-Associated Membranous Nephropathy

    doi: 10.3389/fimmu.2018.03035

    Figure Lengend Snippet: Distribution of PLA 2 R1- and THSD7A-specific IgG subclasses at baseline. (A) Detection of antigen-specific IgG subclasses in the complete study cohort by Western Blot using recombinant proteins. All patients were found to be positive for antigen-specific IgG4-antibodies. Antigen-specific IgG3 antibodies were more common than antigen-specific IgG1, while IgG2 was detected less often. The distribution of (B) PLA 2 R1- and (C) THSD7A-specific IgG subclasses was not different in patients with or without malignancy. Patients with (D) PLA 2 R1- or (E) THSD7A-associated MN were grouped depending on the clinical outcome: negativity of autoantibodies during follow-up, remission of proteinuria, doubling of serum creatinine. IgG1-3 depicts the percentage of patients in whom at least one antigen-specific IgG subclass (IgG1, IgG2, or IgG3) was detected in addition to IgG4.

    Article Snippet: For detection of subclass-specific IgG in Western blot analyses, horseradish peroxidase-conjugated monoclonal anti-human IgG1 (9054-05), IgG2 (9060-05), IgG3 (9210-05), IgG4 (9200-05), and IgG (9040-05) antibodies from mouse were used (IgG1, IgG2, and IgG3 at a 1:5,000 dilution; IgG4 at a 1:10,000 dilution; IgG at a 1:20,000 dilution, all SouthernBiotech, Birmingham, USA), diluted in 5% dry milk in PBS-T.

    Techniques: Proximity Ligation Assay, Western Blot, Recombinant

    GaMD immune serum from WT mice inhibits GaMD-induced renal disease without decreasing other isotypes if injected into GaMD-immunized γ1 - mice by 5d after immunization a . BALB/c γ1 - mice (4 or 8/gp) were injected s.c. with GaMD on day 0 and i.p. with 0.5 ml of pooled serum from GaMD-immunized WT mice (GaMD immune WT serum) or unimmunized WT mice (non-immune serum), starting 4, 5, or 6d after GaMD immunization. Day 7 urine samples were analyzed. ND = none detected. b. BALB/c γ1 - mice (4 or 8/gp) were injected s.c. with GaMD on day 0 and i.p. with 0.5 ml of pooled serum from GaMD-immunized WT mice (GaMD immune WT serum) or unimmunized WT mice (non-immune serum), 5, 6 and 7 d after GaMD immunization. Sera were assayed for total IgG1, IgG2a, IgM and IgG3 on d0 (unimmunized) and 8d after GaMD immunization. ND = none detected.For both a and b , * indicates p

    Journal: Nature

    Article Title: IgG1 protects against renal disease in a mouse model of cryoglobulinemia

    doi: 10.1038/nature13868

    Figure Lengend Snippet: GaMD immune serum from WT mice inhibits GaMD-induced renal disease without decreasing other isotypes if injected into GaMD-immunized γ1 - mice by 5d after immunization a . BALB/c γ1 - mice (4 or 8/gp) were injected s.c. with GaMD on day 0 and i.p. with 0.5 ml of pooled serum from GaMD-immunized WT mice (GaMD immune WT serum) or unimmunized WT mice (non-immune serum), starting 4, 5, or 6d after GaMD immunization. Day 7 urine samples were analyzed. ND = none detected. b. BALB/c γ1 - mice (4 or 8/gp) were injected s.c. with GaMD on day 0 and i.p. with 0.5 ml of pooled serum from GaMD-immunized WT mice (GaMD immune WT serum) or unimmunized WT mice (non-immune serum), 5, 6 and 7 d after GaMD immunization. Sera were assayed for total IgG1, IgG2a, IgM and IgG3 on d0 (unimmunized) and 8d after GaMD immunization. ND = none detected.For both a and b , * indicates p

    Article Snippet: Serum, splenic supernatants and re-suspended cryoglobulin pellets were analyzed for total and GIgG-specific IgG1, IgG2a/c, IgG2b, IgG3, IgE, IgA and IgM content, using standard sandwich ELISA with paired anti-Ig isotype mAbs for each Ig isotype (BD-Pharmingen and eBioscience).

    Techniques: Mouse Assay, Injection

    The development of kidney disease in GaMD-immunized γ1 - mice is independent of IFN-γ, IgG2a, C3 and FcRγ BALB/c WT and γ1 - mice (5/gp) were immunized with GaMD on d0 and injected with 1 mg of either anti-IFN-γ or control mAb on days 0 and 5. a , Total levels of all Ig isotypes were determined in 24 hr culture supernatants of spleen cells harvested on days shown. b , GaMD-immunized γ1 - , γ1 - /FcRγ - , γ1 - /C3 - , C3 - / FcRγ - and γ1 - /C3 - /FcγR - mice (5/gp) had their urine tested for LE and blood on days shown. c , d , BALB/c WT and γ1 - mice (5/gp) were immunized with GaMD on d0 and injected with 1 mg of either anti-IFN-γ or control mAb on days 0 and 5. c , Urine obtained on days indicated was assayed for protein, LE and blood. d , BUN levels were determined prior to and 10 days after GaMD immunization. * p

    Journal: Nature

    Article Title: IgG1 protects against renal disease in a mouse model of cryoglobulinemia

    doi: 10.1038/nature13868

    Figure Lengend Snippet: The development of kidney disease in GaMD-immunized γ1 - mice is independent of IFN-γ, IgG2a, C3 and FcRγ BALB/c WT and γ1 - mice (5/gp) were immunized with GaMD on d0 and injected with 1 mg of either anti-IFN-γ or control mAb on days 0 and 5. a , Total levels of all Ig isotypes were determined in 24 hr culture supernatants of spleen cells harvested on days shown. b , GaMD-immunized γ1 - , γ1 - /FcRγ - , γ1 - /C3 - , C3 - / FcRγ - and γ1 - /C3 - /FcγR - mice (5/gp) had their urine tested for LE and blood on days shown. c , d , BALB/c WT and γ1 - mice (5/gp) were immunized with GaMD on d0 and injected with 1 mg of either anti-IFN-γ or control mAb on days 0 and 5. c , Urine obtained on days indicated was assayed for protein, LE and blood. d , BUN levels were determined prior to and 10 days after GaMD immunization. * p

    Article Snippet: Serum, splenic supernatants and re-suspended cryoglobulin pellets were analyzed for total and GIgG-specific IgG1, IgG2a/c, IgG2b, IgG3, IgE, IgA and IgM content, using standard sandwich ELISA with paired anti-Ig isotype mAbs for each Ig isotype (BD-Pharmingen and eBioscience).

    Techniques: Mouse Assay, Injection

    IgG1 inhibits IgG3-induced cryoglobulin kidney disease independent of complement and FcγRIIB and better than IgG2a and IgG2b a , WT mice (4/gp) were injected i.v. with 4 mg of mouse IgG1, IgG2a, IgG2b, or IgG3 anti-TNP mAb and s.c. with 100 μl of TNP-goat serum on days 0 and 1. Urine LE and blood measured prior to injections and on d1 and d2. b , Urine LE and blood for BALB/c WT and C3 - mice (4/gp) injected i.v. with 4 mg of IgG3 anti-TNP mAb and s.c. with 400 μl of TNP-goat serum on d0 and 1. c , WT and FcγRIIB-deficient (FcγRIIB - ) mice (4/gp) were injected s.c. with 100 μl of TNP-goat serum and i.v. with 4 mg of IgG3 anti-TNP ± 5 mg of IgG1 anti-TNP on d0 and 1. Urinalysis on d0, 1 and 2. d , BALB/c mice were injected i.v. with 4 mg of IgG3 anti-TNP and s.c. with 1.4 mg of TNP-BSA on days 0 and 1. Some mice were also injected with 0.625, 1.25, 2.5, or 5 mg of switch variants of IgG1, IgG2a or IgG2b anti-TNP mAbs on d0 and 1. Urine protein was determined on d0 (not shown), d1 (upper panel) and d2 (lower panel). Results are pooled from a total of 7 experiments. Group size: IgG3 alone: 19 mice; 0.625 mg of IgG1, IgG2a, or IgG2b: 4 mice; 1.25 mg of IgG1, IgG2a or IgG2b: 8 mice; 2.5 mg of IgG1, IgG2a or IgG2b: 6 mice; 5 mg of IgG1, IgG2a or IgG2b: 8 or 9 mice. The significance of differences between treatment groups was determined as described in the legend to Fig. 4f . # p

    Journal: Nature

    Article Title: IgG1 protects against renal disease in a mouse model of cryoglobulinemia

    doi: 10.1038/nature13868

    Figure Lengend Snippet: IgG1 inhibits IgG3-induced cryoglobulin kidney disease independent of complement and FcγRIIB and better than IgG2a and IgG2b a , WT mice (4/gp) were injected i.v. with 4 mg of mouse IgG1, IgG2a, IgG2b, or IgG3 anti-TNP mAb and s.c. with 100 μl of TNP-goat serum on days 0 and 1. Urine LE and blood measured prior to injections and on d1 and d2. b , Urine LE and blood for BALB/c WT and C3 - mice (4/gp) injected i.v. with 4 mg of IgG3 anti-TNP mAb and s.c. with 400 μl of TNP-goat serum on d0 and 1. c , WT and FcγRIIB-deficient (FcγRIIB - ) mice (4/gp) were injected s.c. with 100 μl of TNP-goat serum and i.v. with 4 mg of IgG3 anti-TNP ± 5 mg of IgG1 anti-TNP on d0 and 1. Urinalysis on d0, 1 and 2. d , BALB/c mice were injected i.v. with 4 mg of IgG3 anti-TNP and s.c. with 1.4 mg of TNP-BSA on days 0 and 1. Some mice were also injected with 0.625, 1.25, 2.5, or 5 mg of switch variants of IgG1, IgG2a or IgG2b anti-TNP mAbs on d0 and 1. Urine protein was determined on d0 (not shown), d1 (upper panel) and d2 (lower panel). Results are pooled from a total of 7 experiments. Group size: IgG3 alone: 19 mice; 0.625 mg of IgG1, IgG2a, or IgG2b: 4 mice; 1.25 mg of IgG1, IgG2a or IgG2b: 8 mice; 2.5 mg of IgG1, IgG2a or IgG2b: 6 mice; 5 mg of IgG1, IgG2a or IgG2b: 8 or 9 mice. The significance of differences between treatment groups was determined as described in the legend to Fig. 4f . # p

    Article Snippet: Serum, splenic supernatants and re-suspended cryoglobulin pellets were analyzed for total and GIgG-specific IgG1, IgG2a/c, IgG2b, IgG3, IgE, IgA and IgM content, using standard sandwich ELISA with paired anti-Ig isotype mAbs for each Ig isotype (BD-Pharmingen and eBioscience).

    Techniques: Mouse Assay, Injection

    Concurrent injection of WT mice with IgG3 anti-TNP mAb and TNP-goat serum induces glomerulopathy a , WT mice (4/gp) were injected i.v.with 4 mg of mouse IgG1, IgG2a, IgG2b, or IgG3 anti-TNP mAb and s.c. with 100 μl of TNP-goat serum on days 0 and 1. Urine protein measured prior to injections and on d1 and d2. b , c , WT mice (4/gp) were injected with mouse IgG3 anti-TNP mAb +/- TNP-goat serum as in “a.” Day 2 mouse sera were analyzed for BUN (b). Day 2 kidneys were stained with PAS (c, panel 1: glomerulus from mouse that received only IgG3; panels 2: glomerulus from mouse that received IgG3 + TNP-goat serum). Representative of 3 mice/group. d , Urine protein for BALB/c WT and C3 - mice (4/gp) injected i.v. with 4 mg of IgG3 anti-TNP mAb and s.c. with 400 μl of TNP-goat serum on d0 and 1. * p

    Journal: Nature

    Article Title: IgG1 protects against renal disease in a mouse model of cryoglobulinemia

    doi: 10.1038/nature13868

    Figure Lengend Snippet: Concurrent injection of WT mice with IgG3 anti-TNP mAb and TNP-goat serum induces glomerulopathy a , WT mice (4/gp) were injected i.v.with 4 mg of mouse IgG1, IgG2a, IgG2b, or IgG3 anti-TNP mAb and s.c. with 100 μl of TNP-goat serum on days 0 and 1. Urine protein measured prior to injections and on d1 and d2. b , c , WT mice (4/gp) were injected with mouse IgG3 anti-TNP mAb +/- TNP-goat serum as in “a.” Day 2 mouse sera were analyzed for BUN (b). Day 2 kidneys were stained with PAS (c, panel 1: glomerulus from mouse that received only IgG3; panels 2: glomerulus from mouse that received IgG3 + TNP-goat serum). Representative of 3 mice/group. d , Urine protein for BALB/c WT and C3 - mice (4/gp) injected i.v. with 4 mg of IgG3 anti-TNP mAb and s.c. with 400 μl of TNP-goat serum on d0 and 1. * p

    Article Snippet: Serum, splenic supernatants and re-suspended cryoglobulin pellets were analyzed for total and GIgG-specific IgG1, IgG2a/c, IgG2b, IgG3, IgE, IgA and IgM content, using standard sandwich ELISA with paired anti-Ig isotype mAbs for each Ig isotype (BD-Pharmingen and eBioscience).

    Techniques: Injection, Mouse Assay, Staining

    Glomerulopathy in GaMD-immunized γ1 - mice is complement- and FcRγ -independent and associated with IgG3 cryoglobulinemia a . Serum anti-goat IgG titers in WT and γ1 - mice (4/gp) 8d after GaMD immunization. b , c . Urine protein (b) and BUN (c) of GaMD-immunized γ1 - , γ1 - /FcRγ - , γ1 - /C3 - , C3 - / FcRγ - and γ1 - /C3 - /FcγR - mice (5/gp). d , e . Serum cryoprecipitate protein and Ig isotype concentrations 6-7 d after GaMD immunization of WT and γ1 - mice (7 or 8/gp). Only cryoprecipitates from γ1 - mice contained detectable Ig. f. IgG3 (brown color) in glomerular capillaries (arrows) of γ1 - mice 8d after GaMD (panels 1, low magnification; panel 2, high magnification). * p

    Journal: Nature

    Article Title: IgG1 protects against renal disease in a mouse model of cryoglobulinemia

    doi: 10.1038/nature13868

    Figure Lengend Snippet: Glomerulopathy in GaMD-immunized γ1 - mice is complement- and FcRγ -independent and associated with IgG3 cryoglobulinemia a . Serum anti-goat IgG titers in WT and γ1 - mice (4/gp) 8d after GaMD immunization. b , c . Urine protein (b) and BUN (c) of GaMD-immunized γ1 - , γ1 - /FcRγ - , γ1 - /C3 - , C3 - / FcRγ - and γ1 - /C3 - /FcγR - mice (5/gp). d , e . Serum cryoprecipitate protein and Ig isotype concentrations 6-7 d after GaMD immunization of WT and γ1 - mice (7 or 8/gp). Only cryoprecipitates from γ1 - mice contained detectable Ig. f. IgG3 (brown color) in glomerular capillaries (arrows) of γ1 - mice 8d after GaMD (panels 1, low magnification; panel 2, high magnification). * p

    Article Snippet: Serum, splenic supernatants and re-suspended cryoglobulin pellets were analyzed for total and GIgG-specific IgG1, IgG2a/c, IgG2b, IgG3, IgE, IgA and IgM content, using standard sandwich ELISA with paired anti-Ig isotype mAbs for each Ig isotype (BD-Pharmingen and eBioscience).

    Techniques: Mouse Assay

    Delayed antigen elimination does not account for renal disease in GaMD-immunized γ1 - mice a . BALB/c WT and γ1 - mice (10/gp) were immunized s.c. withGaMD. Sera obtained 5, 6, 7 and 9 days later were evaluated by gel double diffusion for the presence of goat IgG. b-e. BALB/c WT mice (4 or 5/gp) were injected s.c. with a total of 0.2 ml of different mixtures of GaMD and goat anti-KLH antisera. b, c , Mouse sera collected 9d later were assayed for BUN ( b ) and IgG1 anti-goat IgG Ab ( c ). d , Sera obtained 6-13d post-immunization were evaluated by gel double diffusion for the presence of goat IgG. e , Urine samples collected 4-12 days post-immunization were analyzed for protein. * p

    Journal: Nature

    Article Title: IgG1 protects against renal disease in a mouse model of cryoglobulinemia

    doi: 10.1038/nature13868

    Figure Lengend Snippet: Delayed antigen elimination does not account for renal disease in GaMD-immunized γ1 - mice a . BALB/c WT and γ1 - mice (10/gp) were immunized s.c. withGaMD. Sera obtained 5, 6, 7 and 9 days later were evaluated by gel double diffusion for the presence of goat IgG. b-e. BALB/c WT mice (4 or 5/gp) were injected s.c. with a total of 0.2 ml of different mixtures of GaMD and goat anti-KLH antisera. b, c , Mouse sera collected 9d later were assayed for BUN ( b ) and IgG1 anti-goat IgG Ab ( c ). d , Sera obtained 6-13d post-immunization were evaluated by gel double diffusion for the presence of goat IgG. e , Urine samples collected 4-12 days post-immunization were analyzed for protein. * p

    Article Snippet: Serum, splenic supernatants and re-suspended cryoglobulin pellets were analyzed for total and GIgG-specific IgG1, IgG2a/c, IgG2b, IgG3, IgE, IgA and IgM content, using standard sandwich ELISA with paired anti-Ig isotype mAbs for each Ig isotype (BD-Pharmingen and eBioscience).

    Techniques: Mouse Assay, Diffusion-based Assay, Injection

    Ag-specific IgG1 prevents IgG3-mediated glomerulopathy BALB/c γ1 - mice (5/gp) were injected with GaMD on day 0 and/or GaMD-immune or rabbit anti-mouse IgD (RaMD) immune WT serum daily on d4-7. a , b , Urine protein ( a ) and d12 serum albumin and BUN levels ( b ). * p

    Journal: Nature

    Article Title: IgG1 protects against renal disease in a mouse model of cryoglobulinemia

    doi: 10.1038/nature13868

    Figure Lengend Snippet: Ag-specific IgG1 prevents IgG3-mediated glomerulopathy BALB/c γ1 - mice (5/gp) were injected with GaMD on day 0 and/or GaMD-immune or rabbit anti-mouse IgD (RaMD) immune WT serum daily on d4-7. a , b , Urine protein ( a ) and d12 serum albumin and BUN levels ( b ). * p

    Article Snippet: Serum, splenic supernatants and re-suspended cryoglobulin pellets were analyzed for total and GIgG-specific IgG1, IgG2a/c, IgG2b, IgG3, IgE, IgA and IgM content, using standard sandwich ELISA with paired anti-Ig isotype mAbs for each Ig isotype (BD-Pharmingen and eBioscience).

    Techniques: Mouse Assay, Injection

    GaMD immunization of γ1 - mice induces renal dysfunction and glomerular deposition of PAS + material that includes IgG and complement a , WT and γ1 - mice (4/gp) were immunized with GaMD. Urine LE and blood were obtained. b , Representative photomicrographs of glomeruli stained for C3 (top panels) or total mouse IgG (bottom panels) from WT (right panels) and γ1 -/- mice (left panels) 12 d post-GaMD immunization 3 mice/group. c , Deposition of amorphous PAS + material in glomeruli of γ1 - , but not WT begins ∼7 days post GaMD-immunization and leads to glomerular destruction by day 9. Note the scarcity of inflammatory cells in glomeruli. Representative data of 6 mice/group.

    Journal: Nature

    Article Title: IgG1 protects against renal disease in a mouse model of cryoglobulinemia

    doi: 10.1038/nature13868

    Figure Lengend Snippet: GaMD immunization of γ1 - mice induces renal dysfunction and glomerular deposition of PAS + material that includes IgG and complement a , WT and γ1 - mice (4/gp) were immunized with GaMD. Urine LE and blood were obtained. b , Representative photomicrographs of glomeruli stained for C3 (top panels) or total mouse IgG (bottom panels) from WT (right panels) and γ1 -/- mice (left panels) 12 d post-GaMD immunization 3 mice/group. c , Deposition of amorphous PAS + material in glomeruli of γ1 - , but not WT begins ∼7 days post GaMD-immunization and leads to glomerular destruction by day 9. Note the scarcity of inflammatory cells in glomeruli. Representative data of 6 mice/group.

    Article Snippet: Serum, splenic supernatants and re-suspended cryoglobulin pellets were analyzed for total and GIgG-specific IgG1, IgG2a/c, IgG2b, IgG3, IgE, IgA and IgM content, using standard sandwich ELISA with paired anti-Ig isotype mAbs for each Ig isotype (BD-Pharmingen and eBioscience).

    Techniques: Mouse Assay, Staining

    Antigen-specific IgG1 can prevent IgG3 immune complex glomerular deposition BALB/c WT mice were injected i.v.with mouse IgG1 and/or IgG3 anti-TNP mAb with or without s.c.injection of TNP-BSA on days 0 and 1. a. Kidneys were stained with PAS on day 2. Representative micrographs from 3 mice/group are shown. b. Kidney serial sections were stained with PAS or for IgG3 or IgG1 (brown pigment). Representative micrographs from 4 mice/group are shown.

    Journal: Nature

    Article Title: IgG1 protects against renal disease in a mouse model of cryoglobulinemia

    doi: 10.1038/nature13868

    Figure Lengend Snippet: Antigen-specific IgG1 can prevent IgG3 immune complex glomerular deposition BALB/c WT mice were injected i.v.with mouse IgG1 and/or IgG3 anti-TNP mAb with or without s.c.injection of TNP-BSA on days 0 and 1. a. Kidneys were stained with PAS on day 2. Representative micrographs from 3 mice/group are shown. b. Kidney serial sections were stained with PAS or for IgG3 or IgG1 (brown pigment). Representative micrographs from 4 mice/group are shown.

    Article Snippet: Serum, splenic supernatants and re-suspended cryoglobulin pellets were analyzed for total and GIgG-specific IgG1, IgG2a/c, IgG2b, IgG3, IgE, IgA and IgM content, using standard sandwich ELISA with paired anti-Ig isotype mAbs for each Ig isotype (BD-Pharmingen and eBioscience).

    Techniques: Mouse Assay, Injection, Staining

    IgG3 IC persist and accumulate in the glomeruli of GaMD-immunized γ1 - BALB/c WT and γ1 - mice were left untreated or were immunized with GaMD. Kidney sections were stained for mouse IgG1, IgG2a, IgG2b, IgG3 and IgM 8 and 12d later. Representative photomicrographs from 3 GaMD-immunized mice are shown. Insets show magnified views. No staining was observed with sections from unimmunized mice (not shown).

    Journal: Nature

    Article Title: IgG1 protects against renal disease in a mouse model of cryoglobulinemia

    doi: 10.1038/nature13868

    Figure Lengend Snippet: IgG3 IC persist and accumulate in the glomeruli of GaMD-immunized γ1 - BALB/c WT and γ1 - mice were left untreated or were immunized with GaMD. Kidney sections were stained for mouse IgG1, IgG2a, IgG2b, IgG3 and IgM 8 and 12d later. Representative photomicrographs from 3 GaMD-immunized mice are shown. Insets show magnified views. No staining was observed with sections from unimmunized mice (not shown).

    Article Snippet: Serum, splenic supernatants and re-suspended cryoglobulin pellets were analyzed for total and GIgG-specific IgG1, IgG2a/c, IgG2b, IgG3, IgE, IgA and IgM content, using standard sandwich ELISA with paired anti-Ig isotype mAbs for each Ig isotype (BD-Pharmingen and eBioscience).

    Techniques: Mouse Assay, Staining

    GaMD-immunized γ1 +/- mice generate large IgG3 responses but develop mild renal disease BALB/c mice homozygous (γ1 +/+ ), heterozygous (γ1 +/- ) and null (γ1 -/- ) for a functional γ1 allele (6/gp) were injected s.c. with GaMD. a , Sera were titered for goat IgG-specific IgG1, IgG2a and IgG3 0, 8 and 12 days later. Day 0 titers were zero for all Ig isotypes (data not shown). b , Urine samples from the same mice were assayed for protein and leukocyte esterase. ND = none detected.

    Journal: Nature

    Article Title: IgG1 protects against renal disease in a mouse model of cryoglobulinemia

    doi: 10.1038/nature13868

    Figure Lengend Snippet: GaMD-immunized γ1 +/- mice generate large IgG3 responses but develop mild renal disease BALB/c mice homozygous (γ1 +/+ ), heterozygous (γ1 +/- ) and null (γ1 -/- ) for a functional γ1 allele (6/gp) were injected s.c. with GaMD. a , Sera were titered for goat IgG-specific IgG1, IgG2a and IgG3 0, 8 and 12 days later. Day 0 titers were zero for all Ig isotypes (data not shown). b , Urine samples from the same mice were assayed for protein and leukocyte esterase. ND = none detected.

    Article Snippet: Serum, splenic supernatants and re-suspended cryoglobulin pellets were analyzed for total and GIgG-specific IgG1, IgG2a/c, IgG2b, IgG3, IgE, IgA and IgM content, using standard sandwich ELISA with paired anti-Ig isotype mAbs for each Ig isotype (BD-Pharmingen and eBioscience).

    Techniques: Mouse Assay, Functional Assay, Injection

    Lack of C1q affects the level of immunoglobulins IgA (A) , IgG1 (B) , IgG2a (C) , IgG2b (D) , IgG3 (E) , and IgM (F) in infected mice. Whole blood was collected from mice infected with 10 4 Borrelia burgdorferi ) at 7, 10, 21, and 28 days post-infection. Serum level of immunoglobulins was determined by the Bio-Plex system employing the Luminex multianalyte profiling technology. White circles refer to samples from C57BL/6 mice, whereas red squares represent data from C1qα −/− mice ( n = 5; * P

    Journal: Frontiers in Immunology

    Article Title: The Classical Complement Pathway Is Required to Control Borrelia burgdorferi Levels During Experimental Infection

    doi: 10.3389/fimmu.2018.00959

    Figure Lengend Snippet: Lack of C1q affects the level of immunoglobulins IgA (A) , IgG1 (B) , IgG2a (C) , IgG2b (D) , IgG3 (E) , and IgM (F) in infected mice. Whole blood was collected from mice infected with 10 4 Borrelia burgdorferi ) at 7, 10, 21, and 28 days post-infection. Serum level of immunoglobulins was determined by the Bio-Plex system employing the Luminex multianalyte profiling technology. White circles refer to samples from C57BL/6 mice, whereas red squares represent data from C1qα −/− mice ( n = 5; * P

    Article Snippet: To conduct this assay, 96-well microtiter plates were coated with sonicated B. burgdorferi strain B31 at 5 µg/ml in carbonate buffer (pH 9.3), and one empty column (with no antigen) on each plate was reserved for serial dilution of purified mouse IgG1, IgG2a, IgG2b, IgG3, or IgM (eBioscience) to generate a standard curve.

    Techniques: Infection, Mouse Assay, Luminex

    Lack of C1q alters the level of Borrelia -specific antibody subtypes IgM (A) , IgG1 (B) , IgG2a (C) , IgG2b (D) , and IgG3 (E) in infected mice. Whole blood was collected from mice infected with 10 4 Borrelia burgdorferi ) at 7, 10, 21, and 28 days post-infection. Serum level of Borrelia -specific immunoglobulin subtypes was determined by ELISA using total protein lysate derived from B. burgdorferi strain MSK5. White columns refer to C57BL/6 mice, whereas red columns represent data from C1qα −/− mice ( n = 5; * P

    Journal: Frontiers in Immunology

    Article Title: The Classical Complement Pathway Is Required to Control Borrelia burgdorferi Levels During Experimental Infection

    doi: 10.3389/fimmu.2018.00959

    Figure Lengend Snippet: Lack of C1q alters the level of Borrelia -specific antibody subtypes IgM (A) , IgG1 (B) , IgG2a (C) , IgG2b (D) , and IgG3 (E) in infected mice. Whole blood was collected from mice infected with 10 4 Borrelia burgdorferi ) at 7, 10, 21, and 28 days post-infection. Serum level of Borrelia -specific immunoglobulin subtypes was determined by ELISA using total protein lysate derived from B. burgdorferi strain MSK5. White columns refer to C57BL/6 mice, whereas red columns represent data from C1qα −/− mice ( n = 5; * P

    Article Snippet: To conduct this assay, 96-well microtiter plates were coated with sonicated B. burgdorferi strain B31 at 5 µg/ml in carbonate buffer (pH 9.3), and one empty column (with no antigen) on each plate was reserved for serial dilution of purified mouse IgG1, IgG2a, IgG2b, IgG3, or IgM (eBioscience) to generate a standard curve.

    Techniques: Infection, Mouse Assay, Enzyme-linked Immunosorbent Assay, Derivative Assay

    GABA degrading enzyme is expressed in rat kidney. A) RT-PCR results for ABAT in rat kidney cortex. Appropriately-sized bands were detected for ABAT in at least five independent experiments. Rat brain RNA or commercial universal RNA was used as positive control. M: molecular marker. B) Immunoblotting for ABAT in rat kidney cortex. B: brain control, K: kidney cortex. C) Immunostaining for ABAT in rat kidney cortex. Staining with normal IgG instead of primary antibody served as negative control.

    Journal: PLoS ONE

    Article Title: Characteristic Expressions of GABA Receptors and GABA Producing/Transporting Molecules in Rat Kidney

    doi: 10.1371/journal.pone.0105835

    Figure Lengend Snippet: GABA degrading enzyme is expressed in rat kidney. A) RT-PCR results for ABAT in rat kidney cortex. Appropriately-sized bands were detected for ABAT in at least five independent experiments. Rat brain RNA or commercial universal RNA was used as positive control. M: molecular marker. B) Immunoblotting for ABAT in rat kidney cortex. B: brain control, K: kidney cortex. C) Immunostaining for ABAT in rat kidney cortex. Staining with normal IgG instead of primary antibody served as negative control.

    Article Snippet: Normal rabbit IgG (Santa Cruz: sc-2027) and mouse monoclonal IgG1 (Abcam: ab81032) were used as negative control.

    Techniques: Reverse Transcription Polymerase Chain Reaction, Positive Control, Marker, Immunostaining, Staining, Negative Control

    GABA-producing enzymes are expressed in rat kidney. A) RT-PCR results for GAD65 and GAD67 in rat kidney cortex. Appropriately-sized bands were detected for GAD65 and GAD67 in at least five independent experiments. M: molecular marker. B) Immunostaining for GAD65 and GAD67 in rat kidney cortex. Staining with normal IgG instead of primary antibody served as negative control.

    Journal: PLoS ONE

    Article Title: Characteristic Expressions of GABA Receptors and GABA Producing/Transporting Molecules in Rat Kidney

    doi: 10.1371/journal.pone.0105835

    Figure Lengend Snippet: GABA-producing enzymes are expressed in rat kidney. A) RT-PCR results for GAD65 and GAD67 in rat kidney cortex. Appropriately-sized bands were detected for GAD65 and GAD67 in at least five independent experiments. M: molecular marker. B) Immunostaining for GAD65 and GAD67 in rat kidney cortex. Staining with normal IgG instead of primary antibody served as negative control.

    Article Snippet: Normal rabbit IgG (Santa Cruz: sc-2027) and mouse monoclonal IgG1 (Abcam: ab81032) were used as negative control.

    Techniques: Reverse Transcription Polymerase Chain Reaction, Marker, Immunostaining, Staining, Negative Control

    GABA A receptor subunits are expressed in rat kidney. A) RT-PCR results for GABA A receptor subunits in rat (WKY on normal diet) kidney cortex. Appropriately-sized bands were detected for GABA A receptor α1, β3 and π subunits in at least five independent experiments. Rat brain RNA or commercial universal RNA was used as positive control. M: molecular marker. B) Immunoblot for GABA A receptor α1, β3 and π subunits in rat kidney cortex. B: brain control, K: kidney cortex. C) Immunofluorescent examination of GABA A receptor α1, β3 and π subunits in rat kidney cortex. Staining with normal IgG instead of primary antibody served as negative control.

    Journal: PLoS ONE

    Article Title: Characteristic Expressions of GABA Receptors and GABA Producing/Transporting Molecules in Rat Kidney

    doi: 10.1371/journal.pone.0105835

    Figure Lengend Snippet: GABA A receptor subunits are expressed in rat kidney. A) RT-PCR results for GABA A receptor subunits in rat (WKY on normal diet) kidney cortex. Appropriately-sized bands were detected for GABA A receptor α1, β3 and π subunits in at least five independent experiments. Rat brain RNA or commercial universal RNA was used as positive control. M: molecular marker. B) Immunoblot for GABA A receptor α1, β3 and π subunits in rat kidney cortex. B: brain control, K: kidney cortex. C) Immunofluorescent examination of GABA A receptor α1, β3 and π subunits in rat kidney cortex. Staining with normal IgG instead of primary antibody served as negative control.

    Article Snippet: Normal rabbit IgG (Santa Cruz: sc-2027) and mouse monoclonal IgG1 (Abcam: ab81032) were used as negative control.

    Techniques: Reverse Transcription Polymerase Chain Reaction, Positive Control, Marker, Staining, Negative Control

    GABA transporters are expressed in rat kidney. A) RT-PCR results for GAT1, GAT2 and GAT3 in rat kidney cortex. Appropriately-sized bands were detected for GAT2 in at least five independent experiments. Rat brain RNA or commercial universal RNA was used as positive control. M: molecular marker. B) Immunostaining for GAT2 in rat kidney cortex. Staining with normal IgG instead of primary antibody served as negative control.

    Journal: PLoS ONE

    Article Title: Characteristic Expressions of GABA Receptors and GABA Producing/Transporting Molecules in Rat Kidney

    doi: 10.1371/journal.pone.0105835

    Figure Lengend Snippet: GABA transporters are expressed in rat kidney. A) RT-PCR results for GAT1, GAT2 and GAT3 in rat kidney cortex. Appropriately-sized bands were detected for GAT2 in at least five independent experiments. Rat brain RNA or commercial universal RNA was used as positive control. M: molecular marker. B) Immunostaining for GAT2 in rat kidney cortex. Staining with normal IgG instead of primary antibody served as negative control.

    Article Snippet: Normal rabbit IgG (Santa Cruz: sc-2027) and mouse monoclonal IgG1 (Abcam: ab81032) were used as negative control.

    Techniques: Reverse Transcription Polymerase Chain Reaction, Positive Control, Marker, Immunostaining, Staining, Negative Control

    GABA B receptor R1 and R2 subtypes are expressed in rat kidney. A) RT-PCR results for GABA B receptor subtypes in rat kidney cortex. Appropriately-sized bands were detected for GABA B receptor R1 and R2 subtypes in at least five independent experiments. Rat brain RNA or commercial universal RNA was used as positive control. M: molecular marker. B) Immunostaining for GABA B receptor R1 and R2 subtypes in rat kidney cortex. Staining with normal IgG instead of primary antibody served as negative control.

    Journal: PLoS ONE

    Article Title: Characteristic Expressions of GABA Receptors and GABA Producing/Transporting Molecules in Rat Kidney

    doi: 10.1371/journal.pone.0105835

    Figure Lengend Snippet: GABA B receptor R1 and R2 subtypes are expressed in rat kidney. A) RT-PCR results for GABA B receptor subtypes in rat kidney cortex. Appropriately-sized bands were detected for GABA B receptor R1 and R2 subtypes in at least five independent experiments. Rat brain RNA or commercial universal RNA was used as positive control. M: molecular marker. B) Immunostaining for GABA B receptor R1 and R2 subtypes in rat kidney cortex. Staining with normal IgG instead of primary antibody served as negative control.

    Article Snippet: Normal rabbit IgG (Santa Cruz: sc-2027) and mouse monoclonal IgG1 (Abcam: ab81032) were used as negative control.

    Techniques: Reverse Transcription Polymerase Chain Reaction, Positive Control, Marker, Immunostaining, Staining, Negative Control

    C/EBPα preferentially binds and activates the endogenous bcl-2 gene P2 promoter ( a ) Diagram of the human bcl-2 gene. P2 transcription initiates at −58 relative to the initial ATG codon, and P1 initiates at several sites in the vicinity of −1400. The P2, but not the P1, promoter contains a TATAA box (at −88). ( b ) Ba/F3-C/EBPα-ER cells withdrawn from IL-3 were cultured with (+) or without (−) estradiol (E2). The expression of bcl-2 mRNA at the indicated time points was assessed by Northern blot analysis (top). The positions of P1 and P2 bcl-2 transcripts are indicated and RNA loading was assessed by ethidium bromide staining of the 28S and 18S ribosomal RNAs (bottom). ( c ) Ba/F3 MT-C/EBPα, MT-F3901, or parental Ba/F3 cells were cultured with zinc for 16 hours and then withdrawn from IL-3. RNA was extracted 24 hours after IL-3 removal. The levels of P1 and P2 transcripts were analyzed by quantitative RT-PCR and normalized to mS16 expression. Fold-activation relative to the parental Ba/F3 cells is presented. ( d ) Ba/F3 MT-F3901 or MT-C/EBPα cells were treated with zinc for 16 hours and subjected to ChIP using antisera against C/EBPα (α), NF-κB p50, or normal rabbit IgG and primers specific for the P1 or P2 bcl-2 promoters. Data representative of two independent experiments is shown.

    Journal: Leukemia : official journal of the Leukemia Society of America, Leukemia Research Fund, U.K

    Article Title: C/EBP? or C/EBP? oncoproteins regulate the intrinsic and extrinsic apoptotic pathways by direct interaction with NF-?B p50 bound to the bcl-2 and FLIP gene promoters

    doi: 10.1038/leu.2008.297

    Figure Lengend Snippet: C/EBPα preferentially binds and activates the endogenous bcl-2 gene P2 promoter ( a ) Diagram of the human bcl-2 gene. P2 transcription initiates at −58 relative to the initial ATG codon, and P1 initiates at several sites in the vicinity of −1400. The P2, but not the P1, promoter contains a TATAA box (at −88). ( b ) Ba/F3-C/EBPα-ER cells withdrawn from IL-3 were cultured with (+) or without (−) estradiol (E2). The expression of bcl-2 mRNA at the indicated time points was assessed by Northern blot analysis (top). The positions of P1 and P2 bcl-2 transcripts are indicated and RNA loading was assessed by ethidium bromide staining of the 28S and 18S ribosomal RNAs (bottom). ( c ) Ba/F3 MT-C/EBPα, MT-F3901, or parental Ba/F3 cells were cultured with zinc for 16 hours and then withdrawn from IL-3. RNA was extracted 24 hours after IL-3 removal. The levels of P1 and P2 transcripts were analyzed by quantitative RT-PCR and normalized to mS16 expression. Fold-activation relative to the parental Ba/F3 cells is presented. ( d ) Ba/F3 MT-F3901 or MT-C/EBPα cells were treated with zinc for 16 hours and subjected to ChIP using antisera against C/EBPα (α), NF-κB p50, or normal rabbit IgG and primers specific for the P1 or P2 bcl-2 promoters. Data representative of two independent experiments is shown.

    Article Snippet: They were briefly centrifuged, and input was obtained before incubating the resulting supernatant with the antisera against C/EBPα, C/EBPβ, NF-κB p50, NF-κB p65 or rabbit IgG (Santa Cruz) overnight at 4°C with rocking.

    Techniques: Cell Culture, Expressing, Northern Blot, Staining, Quantitative RT-PCR, Activation Assay, Chromatin Immunoprecipitation

    C/EBPα and a C/EBPαLZ oncoprotein depend on NF-κB p50 for binding to the bcl-2 promoter ( a ) Splenocytes from mice with the indicated genotype were exposed to 200 cGy and cultured for 0, 7, or 24 hours. Total cellular proteins extracts were obtained and subjected to Western blotting for bcl-2 and β-tubulin. The ratio of bcl-2:tubulin in each sample is shown. ( b ) Total bone marrow cells extracted from nfkb1 −/− or wild-type control mice were subjected to ChIP using C/EBPα (α) or NF-κB p50 (p50) antisera or IgG control and primers specific for the bcl-2 P2 or neutrophil elastase (NE) promoters. ( c ) RNA isolated from total bone marrow cells from the hind limbs of age, sex, and strain-matched nfkb1 −/− or wild-type (WT) mice were subjected to quantitative RT-PCR analysis of bcl-2 or NE expression, normalized to mS16. Relative mRNA expression between WT and nfkb1 −/− mice is shown, with expression in nfkb1 −/− marrow set to 1. Data from four comparisons are shown. ( d ) F9 cells were transiently co-transfected with 1.5 µg of P2-LUC or its variant harboring clustered point mutations in the −170 κB site (mκB), with 100 ng of CMV, CMV-C/EBPα, or CMV-F3901, and 5 with ng of CMV-β-galactosidase as an internal control. Activation of the wild type and mutant promoters by C/EBPα or F3901 was analyzed 48 after transfection. Fold-activation compared to the empty CMV vector was determined after adjustment for β-galactosidase activity. The mean of four independent experiments is presented. Also shown is the average ratio of P2-LUC:P2-LUCmκB induction by C/EBPα or F3901. The p-values shown compare these induction ratios to the null hypothesis value of 1.0.

    Journal: Leukemia : official journal of the Leukemia Society of America, Leukemia Research Fund, U.K

    Article Title: C/EBP? or C/EBP? oncoproteins regulate the intrinsic and extrinsic apoptotic pathways by direct interaction with NF-?B p50 bound to the bcl-2 and FLIP gene promoters

    doi: 10.1038/leu.2008.297

    Figure Lengend Snippet: C/EBPα and a C/EBPαLZ oncoprotein depend on NF-κB p50 for binding to the bcl-2 promoter ( a ) Splenocytes from mice with the indicated genotype were exposed to 200 cGy and cultured for 0, 7, or 24 hours. Total cellular proteins extracts were obtained and subjected to Western blotting for bcl-2 and β-tubulin. The ratio of bcl-2:tubulin in each sample is shown. ( b ) Total bone marrow cells extracted from nfkb1 −/− or wild-type control mice were subjected to ChIP using C/EBPα (α) or NF-κB p50 (p50) antisera or IgG control and primers specific for the bcl-2 P2 or neutrophil elastase (NE) promoters. ( c ) RNA isolated from total bone marrow cells from the hind limbs of age, sex, and strain-matched nfkb1 −/− or wild-type (WT) mice were subjected to quantitative RT-PCR analysis of bcl-2 or NE expression, normalized to mS16. Relative mRNA expression between WT and nfkb1 −/− mice is shown, with expression in nfkb1 −/− marrow set to 1. Data from four comparisons are shown. ( d ) F9 cells were transiently co-transfected with 1.5 µg of P2-LUC or its variant harboring clustered point mutations in the −170 κB site (mκB), with 100 ng of CMV, CMV-C/EBPα, or CMV-F3901, and 5 with ng of CMV-β-galactosidase as an internal control. Activation of the wild type and mutant promoters by C/EBPα or F3901 was analyzed 48 after transfection. Fold-activation compared to the empty CMV vector was determined after adjustment for β-galactosidase activity. The mean of four independent experiments is presented. Also shown is the average ratio of P2-LUC:P2-LUCmκB induction by C/EBPα or F3901. The p-values shown compare these induction ratios to the null hypothesis value of 1.0.

    Article Snippet: They were briefly centrifuged, and input was obtained before incubating the resulting supernatant with the antisera against C/EBPα, C/EBPβ, NF-κB p50, NF-κB p65 or rabbit IgG (Santa Cruz) overnight at 4°C with rocking.

    Techniques: Binding Assay, Mouse Assay, Cell Culture, Western Blot, Chromatin Immunoprecipitation, Isolation, Quantitative RT-PCR, Expressing, Transfection, Variant Assay, Activation Assay, Mutagenesis, Plasmid Preparation, Activity Assay

    C/EBPα induces FLIP expression and binds its endogenous promoter ( a ) Parental Ba/F3 cells and lines expressing C/EBPα or C/EBPαF3901 from the MT promoter were cultured in zinc chloride for 16 hours and then withdrawn from IL-3. RNA was extracted 16 hours after IL-3 removal. FLIP transcripts were measured using quantitative RT-PCR and expressed as fold-activation compared with parental cells. ( b ) Ba/F3 MT-C/EBPα or MT-C/EBPαF3901 or parental Ba/F3 cells were cultured with zinc for 16 hours and subjected to ChIP using C/EBPα (α) or NF-κB p50 (p50) antisera or control rabbit IgG and primers corresponding to the FLIP promoter. Data representative of two independent experiments is shown. ( c ) Splenocytes from H2K-C/EBPα-Eµ transgenic (αTG) or nfkb1 −/−;αTG mice were subjected to ChIP using C/EBPα antiserum. Shown is the ratio of signal detected in the immunoprecipitate compared with input in three repetitions. ( d ) Single cell suspensions of splenocytes were obtained from mice with the indicated genotypes, stimulated with LPS for 30 hours and cultured with 200 ng/mL of soluble FasL for 16 hours. The cells were then stained with APC-Annexin V and PI. Shown are the percent of cells that were Annexin V-negative, all of which excluded PI (mean and SD from three experiments).

    Journal: Leukemia : official journal of the Leukemia Society of America, Leukemia Research Fund, U.K

    Article Title: C/EBP? or C/EBP? oncoproteins regulate the intrinsic and extrinsic apoptotic pathways by direct interaction with NF-?B p50 bound to the bcl-2 and FLIP gene promoters

    doi: 10.1038/leu.2008.297

    Figure Lengend Snippet: C/EBPα induces FLIP expression and binds its endogenous promoter ( a ) Parental Ba/F3 cells and lines expressing C/EBPα or C/EBPαF3901 from the MT promoter were cultured in zinc chloride for 16 hours and then withdrawn from IL-3. RNA was extracted 16 hours after IL-3 removal. FLIP transcripts were measured using quantitative RT-PCR and expressed as fold-activation compared with parental cells. ( b ) Ba/F3 MT-C/EBPα or MT-C/EBPαF3901 or parental Ba/F3 cells were cultured with zinc for 16 hours and subjected to ChIP using C/EBPα (α) or NF-κB p50 (p50) antisera or control rabbit IgG and primers corresponding to the FLIP promoter. Data representative of two independent experiments is shown. ( c ) Splenocytes from H2K-C/EBPα-Eµ transgenic (αTG) or nfkb1 −/−;αTG mice were subjected to ChIP using C/EBPα antiserum. Shown is the ratio of signal detected in the immunoprecipitate compared with input in three repetitions. ( d ) Single cell suspensions of splenocytes were obtained from mice with the indicated genotypes, stimulated with LPS for 30 hours and cultured with 200 ng/mL of soluble FasL for 16 hours. The cells were then stained with APC-Annexin V and PI. Shown are the percent of cells that were Annexin V-negative, all of which excluded PI (mean and SD from three experiments).

    Article Snippet: They were briefly centrifuged, and input was obtained before incubating the resulting supernatant with the antisera against C/EBPα, C/EBPβ, NF-κB p50, NF-κB p65 or rabbit IgG (Santa Cruz) overnight at 4°C with rocking.

    Techniques: Expressing, Cell Culture, Quantitative RT-PCR, Activation Assay, Chromatin Immunoprecipitation, Transgenic Assay, Mouse Assay, Staining

    The C/EBPα basic region mediates its interaction with NF-κB p50 or p65 ( a ) 293T cells in 100 mm dishes were cotransfected with 2 µg CMV-C/EBPαbZIP and 2 µg of either CMV-NFκB p50 or CMV-NF-κB p65. Two days later, cells extracts were immunoprecipitated with p50 or p65 rabbit antisera (Ab) or rabbit IgG control (Ig) and immunoblotted with rabbit antiserum raised against the C/EBPα COOH-terminal sequence. ( b ) Extracts from cells transfected with CMV-C/EBPα or CMV-C/EBPαGZ and either CMV-NF-κB p50 or CMV-NF-κB p65 were immunoprecipitated with C/EBPα antiserum or rabbit IgG and immunoblotted with monoclonal p50 or p65 antibodies, as indicated (right and left panels). Extracts from cells transfected with p50 or p65 alone were assayed similarly as additional controls (center panel).

    Journal: Leukemia : official journal of the Leukemia Society of America, Leukemia Research Fund, U.K

    Article Title: C/EBP? or C/EBP? oncoproteins regulate the intrinsic and extrinsic apoptotic pathways by direct interaction with NF-?B p50 bound to the bcl-2 and FLIP gene promoters

    doi: 10.1038/leu.2008.297

    Figure Lengend Snippet: The C/EBPα basic region mediates its interaction with NF-κB p50 or p65 ( a ) 293T cells in 100 mm dishes were cotransfected with 2 µg CMV-C/EBPαbZIP and 2 µg of either CMV-NFκB p50 or CMV-NF-κB p65. Two days later, cells extracts were immunoprecipitated with p50 or p65 rabbit antisera (Ab) or rabbit IgG control (Ig) and immunoblotted with rabbit antiserum raised against the C/EBPα COOH-terminal sequence. ( b ) Extracts from cells transfected with CMV-C/EBPα or CMV-C/EBPαGZ and either CMV-NF-κB p50 or CMV-NF-κB p65 were immunoprecipitated with C/EBPα antiserum or rabbit IgG and immunoblotted with monoclonal p50 or p65 antibodies, as indicated (right and left panels). Extracts from cells transfected with p50 or p65 alone were assayed similarly as additional controls (center panel).

    Article Snippet: They were briefly centrifuged, and input was obtained before incubating the resulting supernatant with the antisera against C/EBPα, C/EBPβ, NF-κB p50, NF-κB p65 or rabbit IgG (Santa Cruz) overnight at 4°C with rocking.

    Techniques: Immunoprecipitation, Sequencing, Transfection

    C/EBPα and a C/EBPαLZ oncoprotein bind the endogenous bcl-2 promoter ( a ) HF-1 cells were subjected to ChIP analysis using antisera against C/EBPα (α), C/EBPβ (β), NF-κB p50, NF-κB p65, or IgG control. PCR products were subjected to agarose gel electropheresis and visualized using ethidium bromide. In – input, 1% of DNA used for ChIP. ( b ) Total cellular proteins from Ba/F3 lines expressing the indicated MT-C/EBPα isoform or from parental cells were subjected to Western blotting using C/EBPα and β-tubulin antisera. ( c ) Ba/F3 cells expressing MT-C/EBPα or MT-F3901, or parental Ba/F3 cells, were cultured with zinc chloride for 16 hours and subjected to ChIP analysis. After immunoprecipitation with antiserum against C/EBPα (α) or rabbit IgG, the precipitated DNAs were subjected to PCR for the bcl-2 or neutrophil elastase (NE) promoters, as indicated. Data representative of 3 independent experiments is shown.

    Journal: Leukemia : official journal of the Leukemia Society of America, Leukemia Research Fund, U.K

    Article Title: C/EBP? or C/EBP? oncoproteins regulate the intrinsic and extrinsic apoptotic pathways by direct interaction with NF-?B p50 bound to the bcl-2 and FLIP gene promoters

    doi: 10.1038/leu.2008.297

    Figure Lengend Snippet: C/EBPα and a C/EBPαLZ oncoprotein bind the endogenous bcl-2 promoter ( a ) HF-1 cells were subjected to ChIP analysis using antisera against C/EBPα (α), C/EBPβ (β), NF-κB p50, NF-κB p65, or IgG control. PCR products were subjected to agarose gel electropheresis and visualized using ethidium bromide. In – input, 1% of DNA used for ChIP. ( b ) Total cellular proteins from Ba/F3 lines expressing the indicated MT-C/EBPα isoform or from parental cells were subjected to Western blotting using C/EBPα and β-tubulin antisera. ( c ) Ba/F3 cells expressing MT-C/EBPα or MT-F3901, or parental Ba/F3 cells, were cultured with zinc chloride for 16 hours and subjected to ChIP analysis. After immunoprecipitation with antiserum against C/EBPα (α) or rabbit IgG, the precipitated DNAs were subjected to PCR for the bcl-2 or neutrophil elastase (NE) promoters, as indicated. Data representative of 3 independent experiments is shown.

    Article Snippet: They were briefly centrifuged, and input was obtained before incubating the resulting supernatant with the antisera against C/EBPα, C/EBPβ, NF-κB p50, NF-κB p65 or rabbit IgG (Santa Cruz) overnight at 4°C with rocking.

    Techniques: Chromatin Immunoprecipitation, Polymerase Chain Reaction, Agarose Gel Electrophoresis, Expressing, Western Blot, Cell Culture, Immunoprecipitation