mouse complement c3  (Valiant Co Ltd)

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

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

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