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antiserum/complement mixture (1:20 dilution of the serum and 20% of mouse complement  (Thermo Fisher)


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    Thermo Fisher antiserum/complement mixture (1:20 dilution of the serum and 20% of mouse complement
    Antiserum/Complement Mixture (1:20 Dilution Of The Serum And 20% Of Mouse Complement, 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
    https://www.bioz.com/product/mouse+serum+complement/pm39370617-339-12-26
    Average 90 stars, based on 1 article reviews
    antiserum/complement mixture (1:20 dilution of the serum and 20% of mouse complement - by Bioz Stars, 2026-11
    90/100 stars

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    Article Title: Mechanism of allorecognition and skin graft rejection in CD28 and CD40 ligand double-deficient mice
    Article Snippet: Background.. It has been shown that simultaneous blockade of CD28and CD40-mediated costimulatory signals significantly prolongs allograft survival.. Although these results led to an expectation of the establishment of specific immunotolerant therapy for organ transplantation, it became evident that these treatments rarely resulted in indefinite allograft survival.

    Article Title: Immunogenic Sulfated l-Idose Homo Oligosaccharides Elicit Neutralizing Antibody against Native Heparan Sulfate with Biomarker and Therapeutic Possibilities.
    Article Snippet: Heparan sulfate (HS) is a non-immunogenic antigen, and developing antibodies against specific sulfated patterns in HS poses significant challenges.. Herein, we employed an innovative immunization strategy that exploits the molecular mimicry of HS to generate antibodies against HS sequences.. Mice were immunized with synthetic sulfated oligo-L-idose (ID49) that mimics optimum 67% of the conserved structure of HS ligands.

    Article Title: Adiponectin reduces immune checkpoint inhibitor-induced inflammation without blocking anti-tumor immunity.
    Article Snippet: Isolation of immune cell populations Immune cells were isolated for gene expression analysis from bonemarrow using Neutrophil Isolation Kit orMonocyte Isolation Kit, or from the spleen using NK cell Isolation Kit, Pan B-cell Isolation Kit or Pan T cell Isolation Kit II (all for mouse; Miltenyi).

    Extraction:

    Article Title: Differential Interactions of Serum and Bronchoalveolar Lavage Fluid Complement Proteins with Conidia of Airborne Fungal Pathogen Aspergillus fumigatus
    Article Snippet: Anti-MAC labelling was 455 performed using anti-human C5b-9 antibody (mouse monoclonal, clone aE11; Abcam) and 456 secondary FITC-conjugated anti-mouse IgG (Sigma-Aldrich). .. 457 on June 23, 2020 at M acquarie U niversity Library http://iai.asm .org/ D ow nloaded from 458 Conidial opsonization, RodAp extraction using hydrofluoric acid and Western-blot analyses 459 As described above, conidia were opsonized with pooled serum (diluted to 20% in PBS) for 30 460 min, washed five-times with PBS and subjected to RodAp extraction using hydrofluoric acid 461 (HF) (1), but only for 6 h. Extracted samples were subjected to SDS-PAGE on 12% gel and 462 revealed Coomassie-brilliant blue staining or Western-blot by separating proteins on 15% gel 463 and using mouse polyclonal anti-RodAp antibodies (62), anti-C3 (LF-MA0132, mouse 464 monoclonal, clone 28A1, targets C3b; ThermoFisher Scientific) or anti-C3b antibodies [mouse 465 monoclonal, MA1-40155; Invitrogen; targets iC3b-part, checked using human serum as the 466 positive control and C3-deficient serum as the negative control and we cross-checked it with 467 human serum and C3-depleted serum (CompTech, Complement Technology Inc., 468 Catalog#A314)] as the primary antibodies, and peroxidase conjugated anti-mouse IgG as the 469 secondary antibody (1:1000 dilution; Sigma-Aldrich). .. The blot-detection was performed using 470 ECL-chemiluminescence kit (Amersham, GE Healthcare Life Sciences).

    Western Blot:

    Article Title: Differential Interactions of Serum and Bronchoalveolar Lavage Fluid Complement Proteins with Conidia of Airborne Fungal Pathogen Aspergillus fumigatus
    Article Snippet: Anti-MAC labelling was 455 performed using anti-human C5b-9 antibody (mouse monoclonal, clone aE11; Abcam) and 456 secondary FITC-conjugated anti-mouse IgG (Sigma-Aldrich). .. 457 on June 23, 2020 at M acquarie U niversity Library http://iai.asm .org/ D ow nloaded from 458 Conidial opsonization, RodAp extraction using hydrofluoric acid and Western-blot analyses 459 As described above, conidia were opsonized with pooled serum (diluted to 20% in PBS) for 30 460 min, washed five-times with PBS and subjected to RodAp extraction using hydrofluoric acid 461 (HF) (1), but only for 6 h. Extracted samples were subjected to SDS-PAGE on 12% gel and 462 revealed Coomassie-brilliant blue staining or Western-blot by separating proteins on 15% gel 463 and using mouse polyclonal anti-RodAp antibodies (62), anti-C3 (LF-MA0132, mouse 464 monoclonal, clone 28A1, targets C3b; ThermoFisher Scientific) or anti-C3b antibodies [mouse 465 monoclonal, MA1-40155; Invitrogen; targets iC3b-part, checked using human serum as the 466 positive control and C3-deficient serum as the negative control and we cross-checked it with 467 human serum and C3-depleted serum (CompTech, Complement Technology Inc., 468 Catalog#A314)] as the primary antibodies, and peroxidase conjugated anti-mouse IgG as the 469 secondary antibody (1:1000 dilution; Sigma-Aldrich). .. The blot-detection was performed using 470 ECL-chemiluminescence kit (Amersham, GE Healthcare Life Sciences).

    SDS Page:

    Article Title: Differential Interactions of Serum and Bronchoalveolar Lavage Fluid Complement Proteins with Conidia of Airborne Fungal Pathogen Aspergillus fumigatus
    Article Snippet: Anti-MAC labelling was 455 performed using anti-human C5b-9 antibody (mouse monoclonal, clone aE11; Abcam) and 456 secondary FITC-conjugated anti-mouse IgG (Sigma-Aldrich). .. 457 on June 23, 2020 at M acquarie U niversity Library http://iai.asm .org/ D ow nloaded from 458 Conidial opsonization, RodAp extraction using hydrofluoric acid and Western-blot analyses 459 As described above, conidia were opsonized with pooled serum (diluted to 20% in PBS) for 30 460 min, washed five-times with PBS and subjected to RodAp extraction using hydrofluoric acid 461 (HF) (1), but only for 6 h. Extracted samples were subjected to SDS-PAGE on 12% gel and 462 revealed Coomassie-brilliant blue staining or Western-blot by separating proteins on 15% gel 463 and using mouse polyclonal anti-RodAp antibodies (62), anti-C3 (LF-MA0132, mouse 464 monoclonal, clone 28A1, targets C3b; ThermoFisher Scientific) or anti-C3b antibodies [mouse 465 monoclonal, MA1-40155; Invitrogen; targets iC3b-part, checked using human serum as the 466 positive control and C3-deficient serum as the negative control and we cross-checked it with 467 human serum and C3-depleted serum (CompTech, Complement Technology Inc., 468 Catalog#A314)] as the primary antibodies, and peroxidase conjugated anti-mouse IgG as the 469 secondary antibody (1:1000 dilution; Sigma-Aldrich). .. The blot-detection was performed using 470 ECL-chemiluminescence kit (Amersham, GE Healthcare Life Sciences).

    Staining:

    Article Title: Differential Interactions of Serum and Bronchoalveolar Lavage Fluid Complement Proteins with Conidia of Airborne Fungal Pathogen Aspergillus fumigatus
    Article Snippet: Anti-MAC labelling was 455 performed using anti-human C5b-9 antibody (mouse monoclonal, clone aE11; Abcam) and 456 secondary FITC-conjugated anti-mouse IgG (Sigma-Aldrich). .. 457 on June 23, 2020 at M acquarie U niversity Library http://iai.asm .org/ D ow nloaded from 458 Conidial opsonization, RodAp extraction using hydrofluoric acid and Western-blot analyses 459 As described above, conidia were opsonized with pooled serum (diluted to 20% in PBS) for 30 460 min, washed five-times with PBS and subjected to RodAp extraction using hydrofluoric acid 461 (HF) (1), but only for 6 h. Extracted samples were subjected to SDS-PAGE on 12% gel and 462 revealed Coomassie-brilliant blue staining or Western-blot by separating proteins on 15% gel 463 and using mouse polyclonal anti-RodAp antibodies (62), anti-C3 (LF-MA0132, mouse 464 monoclonal, clone 28A1, targets C3b; ThermoFisher Scientific) or anti-C3b antibodies [mouse 465 monoclonal, MA1-40155; Invitrogen; targets iC3b-part, checked using human serum as the 466 positive control and C3-deficient serum as the negative control and we cross-checked it with 467 human serum and C3-depleted serum (CompTech, Complement Technology Inc., 468 Catalog#A314)] as the primary antibodies, and peroxidase conjugated anti-mouse IgG as the 469 secondary antibody (1:1000 dilution; Sigma-Aldrich). .. The blot-detection was performed using 470 ECL-chemiluminescence kit (Amersham, GE Healthcare Life Sciences).

    Positive Control:

    Article Title: Differential Interactions of Serum and Bronchoalveolar Lavage Fluid Complement Proteins with Conidia of Airborne Fungal Pathogen Aspergillus fumigatus
    Article Snippet: Anti-MAC labelling was 455 performed using anti-human C5b-9 antibody (mouse monoclonal, clone aE11; Abcam) and 456 secondary FITC-conjugated anti-mouse IgG (Sigma-Aldrich). .. 457 on June 23, 2020 at M acquarie U niversity Library http://iai.asm .org/ D ow nloaded from 458 Conidial opsonization, RodAp extraction using hydrofluoric acid and Western-blot analyses 459 As described above, conidia were opsonized with pooled serum (diluted to 20% in PBS) for 30 460 min, washed five-times with PBS and subjected to RodAp extraction using hydrofluoric acid 461 (HF) (1), but only for 6 h. Extracted samples were subjected to SDS-PAGE on 12% gel and 462 revealed Coomassie-brilliant blue staining or Western-blot by separating proteins on 15% gel 463 and using mouse polyclonal anti-RodAp antibodies (62), anti-C3 (LF-MA0132, mouse 464 monoclonal, clone 28A1, targets C3b; ThermoFisher Scientific) or anti-C3b antibodies [mouse 465 monoclonal, MA1-40155; Invitrogen; targets iC3b-part, checked using human serum as the 466 positive control and C3-deficient serum as the negative control and we cross-checked it with 467 human serum and C3-depleted serum (CompTech, Complement Technology Inc., 468 Catalog#A314)] as the primary antibodies, and peroxidase conjugated anti-mouse IgG as the 469 secondary antibody (1:1000 dilution; Sigma-Aldrich). .. The blot-detection was performed using 470 ECL-chemiluminescence kit (Amersham, GE Healthcare Life Sciences).

    Negative Control:

    Article Title: Differential Interactions of Serum and Bronchoalveolar Lavage Fluid Complement Proteins with Conidia of Airborne Fungal Pathogen Aspergillus fumigatus
    Article Snippet: Anti-MAC labelling was 455 performed using anti-human C5b-9 antibody (mouse monoclonal, clone aE11; Abcam) and 456 secondary FITC-conjugated anti-mouse IgG (Sigma-Aldrich). .. 457 on June 23, 2020 at M acquarie U niversity Library http://iai.asm .org/ D ow nloaded from 458 Conidial opsonization, RodAp extraction using hydrofluoric acid and Western-blot analyses 459 As described above, conidia were opsonized with pooled serum (diluted to 20% in PBS) for 30 460 min, washed five-times with PBS and subjected to RodAp extraction using hydrofluoric acid 461 (HF) (1), but only for 6 h. Extracted samples were subjected to SDS-PAGE on 12% gel and 462 revealed Coomassie-brilliant blue staining or Western-blot by separating proteins on 15% gel 463 and using mouse polyclonal anti-RodAp antibodies (62), anti-C3 (LF-MA0132, mouse 464 monoclonal, clone 28A1, targets C3b; ThermoFisher Scientific) or anti-C3b antibodies [mouse 465 monoclonal, MA1-40155; Invitrogen; targets iC3b-part, checked using human serum as the 466 positive control and C3-deficient serum as the negative control and we cross-checked it with 467 human serum and C3-depleted serum (CompTech, Complement Technology Inc., 468 Catalog#A314)] as the primary antibodies, and peroxidase conjugated anti-mouse IgG as the 469 secondary antibody (1:1000 dilution; Sigma-Aldrich). .. The blot-detection was performed using 470 ECL-chemiluminescence kit (Amersham, GE Healthcare Life Sciences).



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    A-C: Protein ELISA measuring reactivity of TbpB, C-lobe, and LCL-immune sera against full-length TbpB, TbpB C-lobe, and TbpB N-lobe, respectively. D: IgG titre elicited by the different immunogens towards heat inactivated Nme M982. A-D: N=10-15 per group; squares represent pre-challenge sera from C57BL/6 animals from the sepsis study presented in ; circles represent terminal serum from hCEACAM1 FvB mice from the colonization study presented in , each symbol represents serum from one animal tested in duplicate. E: Serum bactericidal activity (SBA) titre against iron-starved Nme M982. N=19-25 per group, including terminal hCEACAM1 FvB serum from , plus additional terminal serum from immunized hCEACAM1 FvB from pilot studies. SBA titre (>50% killing) reached for 19/25 (76%) TbpB, 8/19 (42%) C-lobe, 14/22 (64%) LCL and 3/21 (14%) Alum samples. Dotted lines represent lowest dilution tested. Line at median for each group. One-way ANOVA with Tukey’s post-hoc test comparing each group to every other group performed using GraphPad Prism 10.2.0. For D, E: Statistics performed on Log2 transformed data. ns, not significant; **, p<0.01; ***, p<0.001; ****, p<0.0001. F: Meningococcal growth inhibition assay. Iron-starved M982 strain was grown in the presence of hTf and pooled heat-inactivated serum from TbpB, C-lobe and LCL immunized animals from . Change in absorbance (OD600) relative to the initial time point is graphed; error bars depict standard deviation of two technical replicates.

    Journal: bioRxiv

    Article Title: Rationally designed minimized TbpB confers broad protection against meningococcal infection

    doi: 10.1101/2025.10.30.685603

    Figure Lengend Snippet: A-C: Protein ELISA measuring reactivity of TbpB, C-lobe, and LCL-immune sera against full-length TbpB, TbpB C-lobe, and TbpB N-lobe, respectively. D: IgG titre elicited by the different immunogens towards heat inactivated Nme M982. A-D: N=10-15 per group; squares represent pre-challenge sera from C57BL/6 animals from the sepsis study presented in ; circles represent terminal serum from hCEACAM1 FvB mice from the colonization study presented in , each symbol represents serum from one animal tested in duplicate. E: Serum bactericidal activity (SBA) titre against iron-starved Nme M982. N=19-25 per group, including terminal hCEACAM1 FvB serum from , plus additional terminal serum from immunized hCEACAM1 FvB from pilot studies. SBA titre (>50% killing) reached for 19/25 (76%) TbpB, 8/19 (42%) C-lobe, 14/22 (64%) LCL and 3/21 (14%) Alum samples. Dotted lines represent lowest dilution tested. Line at median for each group. One-way ANOVA with Tukey’s post-hoc test comparing each group to every other group performed using GraphPad Prism 10.2.0. For D, E: Statistics performed on Log2 transformed data. ns, not significant; **, p<0.01; ***, p<0.001; ****, p<0.0001. F: Meningococcal growth inhibition assay. Iron-starved M982 strain was grown in the presence of hTf and pooled heat-inactivated serum from TbpB, C-lobe and LCL immunized animals from . Change in absorbance (OD600) relative to the initial time point is graphed; error bars depict standard deviation of two technical replicates.

    Article Snippet: Assay was set up in 40 μL total volume using ∼500 bacteria suspended in PBS++, 10% baby rabbit complement (Cedarlane, CL3441-S100) and 2-fold serial dilutions of heat inactivated terminal mouse serum.

    Techniques: Enzyme-linked Immunosorbent Assay, Activity Assay, Transformation Assay, Growth Inhibition Assay, Standard Deviation

    A: Phylogenetic tree depicting Neisserial TbpB diversity. The location of M982 TbpB is highlighted, along with 16 additional TbpBs spanning various clusters used for cross-reactivity analysis. B: Heat map depicting IgG cross-reactivity against a broad panel of TbpB protein variants by ELISA. Optical density (OD) readings denoted by the colour gradient. Background noise from no-protein control subtracted. Each row is a different TbpB protein capture, with the phylogenetic cluster indicated to the left. Each column is a different mouse sample, with the immunizing antigens indicated above. Numbered labels starting with A are pre-challenge serum from the immunized C67BL/6 sepsis cohort; B are terminal serum from the immunized hCEACAM1 FvB colonization cohort. C: Grouped bar graph summarizing cross-reactive antibody data from B. Bars represent mean, error bars represent standard error, each circle represents serum from a single mouse (N=8 for immune sera, N=3 for Alum). hTf-HRP signal quantifying properly folded TbpB capture is depicted in the inset, mean ± standard deviation. D: Serum reactivity against representative Nme and Ngo strains grown under iron limitation using inactivated whole bacterial ELISA. TbpB cluster indicated below the x-axis. Background noise from no serum control subtracted. Bars represent mean, error bars represent standard error, each circle represents serum from a single mouse, N=5-8/group. Inset depicts hTf-binding as an indicator of TbpAB receptor expression on bacteria used for coating ELISA plates, mean ± standard deviation. For C and D, two-way ANOVA with Dunnett’s post-hoc test comparing each group to Alum control performed using GraphPad Prism 10.2.0. Only p-values <0.05 shown. *, p<0.05; **, p<0.01; ***, p<0.001, ****, p<0.0001.

    Journal: bioRxiv

    Article Title: Rationally designed minimized TbpB confers broad protection against meningococcal infection

    doi: 10.1101/2025.10.30.685603

    Figure Lengend Snippet: A: Phylogenetic tree depicting Neisserial TbpB diversity. The location of M982 TbpB is highlighted, along with 16 additional TbpBs spanning various clusters used for cross-reactivity analysis. B: Heat map depicting IgG cross-reactivity against a broad panel of TbpB protein variants by ELISA. Optical density (OD) readings denoted by the colour gradient. Background noise from no-protein control subtracted. Each row is a different TbpB protein capture, with the phylogenetic cluster indicated to the left. Each column is a different mouse sample, with the immunizing antigens indicated above. Numbered labels starting with A are pre-challenge serum from the immunized C67BL/6 sepsis cohort; B are terminal serum from the immunized hCEACAM1 FvB colonization cohort. C: Grouped bar graph summarizing cross-reactive antibody data from B. Bars represent mean, error bars represent standard error, each circle represents serum from a single mouse (N=8 for immune sera, N=3 for Alum). hTf-HRP signal quantifying properly folded TbpB capture is depicted in the inset, mean ± standard deviation. D: Serum reactivity against representative Nme and Ngo strains grown under iron limitation using inactivated whole bacterial ELISA. TbpB cluster indicated below the x-axis. Background noise from no serum control subtracted. Bars represent mean, error bars represent standard error, each circle represents serum from a single mouse, N=5-8/group. Inset depicts hTf-binding as an indicator of TbpAB receptor expression on bacteria used for coating ELISA plates, mean ± standard deviation. For C and D, two-way ANOVA with Dunnett’s post-hoc test comparing each group to Alum control performed using GraphPad Prism 10.2.0. Only p-values <0.05 shown. *, p<0.05; **, p<0.01; ***, p<0.001, ****, p<0.0001.

    Article Snippet: Assay was set up in 40 μL total volume using ∼500 bacteria suspended in PBS++, 10% baby rabbit complement (Cedarlane, CL3441-S100) and 2-fold serial dilutions of heat inactivated terminal mouse serum.

    Techniques: Enzyme-linked Immunosorbent Assay, Control, Standard Deviation, Binding Assay, Expressing, Bacteria

    AT1a receptor loss alleviated denervation-induced muscle atrophy independently of C1q/Wnt/β-catenin signaling in gastrocnemius muscle. ( a , b ) Gene expressions of C1qa ( a ) and Axin2 ( b ) were evaluated by RT-qPCR. Expression levels of these transcripts in each sample were normalized to 18S rRNA expression levels and were shown as the fold increase or decrease in mRNA expression in the Den group compared with the Cont group. ( c ) Serum C1q concentration was measured by ELISA. The graphs show the fold increase or decrease in serum concentration levels in the Den group compared with the Cont group. AT1a +/+ -Cont group, n = 6; AT1a +/+ -Den group at 7 days postdenervation, n = 7; AT1a +/+ -Den group at 21 days postdenervation, n = 7; AT1a −/− -Cont group, n = 5; AT1a −/− -Den group at 7 days postdenervation, n = 6; AT1a −/− -Den group at 21 days postdenervation, n = 6. Values are means ± SE.

    Journal: Scientific Reports

    Article Title: Angiotensin II type 1a receptor deficiency alleviates muscle atrophy after denervation

    doi: 10.1038/s41598-023-27737-7

    Figure Lengend Snippet: AT1a receptor loss alleviated denervation-induced muscle atrophy independently of C1q/Wnt/β-catenin signaling in gastrocnemius muscle. ( a , b ) Gene expressions of C1qa ( a ) and Axin2 ( b ) were evaluated by RT-qPCR. Expression levels of these transcripts in each sample were normalized to 18S rRNA expression levels and were shown as the fold increase or decrease in mRNA expression in the Den group compared with the Cont group. ( c ) Serum C1q concentration was measured by ELISA. The graphs show the fold increase or decrease in serum concentration levels in the Den group compared with the Cont group. AT1a +/+ -Cont group, n = 6; AT1a +/+ -Den group at 7 days postdenervation, n = 7; AT1a +/+ -Den group at 21 days postdenervation, n = 7; AT1a −/− -Cont group, n = 5; AT1a −/− -Den group at 7 days postdenervation, n = 6; AT1a −/− -Den group at 21 days postdenervation, n = 6. Values are means ± SE.

    Article Snippet: Serum complement C1q concentration was measured using a C1q, mouse, ELISA kit (Hycult Biotech, Uden, The Netherlands), which was the same as the kit used in the previous study showing the beneficial effects of Ang II receptor blockade against muscle injury .

    Techniques: Quantitative RT-PCR, Expressing, Concentration Assay, Enzyme-linked Immunosorbent Assay