hycult elisa kit  (Hycult Biotech)

Journal: Journal of Neuroinflammation

Article Title: Systemic inflammation triggers local complement production in the mouse retina and RPE

doi: 10.1186/s12974-026-03737-y

Figure Lengend Snippet: Analysis of C3 expression through ELISA, qPCR and IHC. C3 expression in the serum of control group (t0, saline treated mice), as well as 6, 24 and 72 h after LPS injection ( A ). mRNA expression of C3 24 h post-LPS injection in the retina ( B ) and RPE ( C ). Immunohistochemistry of C3 expression 24 h after systemic LPS injection ( D - G ). Representative retina sections of control group (CTRL, saline treated mice, D - E ) and treated animals 24 h post-LPS injection ( F - G ), stained with anti-C3 antibody. Yellow arrowheads indicate the main localization of the signal (RPE in CTRL, and RPE and ganglion cell layer in the LPS 24 h group). Data are presented as mean ± SEM (experimental n ≥ 4). * P < 0.05, ** P < 0.01, *** P < 0.001 (One-Way ANOVA with Tukey’s multiple comparison test or Unpaired T-test). Abbreviations: GCL: ganglion cell layer; INL: inner nuclear layer; ONL: outer nuclear layer; RPE: retinal pigment epithelium. Scale bar: 50 μm

Article Snippet: C3 serum levels were measured using the Hycult ELISA kit (HK2002, Hycult Biotechnologies, Uden, NL) according to the manufacturer’s instructions.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Control, Saline, Injection, Immunohistochemistry, Staining, Comparison

Journal: Nature Communications

Article Title: Radical footprinting in mammalian whole blood

doi: 10.1038/s41467-026-68982-4

Figure Lengend Snippet: Conformational changes from C3, containing 13 domains ( a ) to C3b ( b ) following complement activation. ANA anaphylatoxin domain, CUB complement C1r/C1s, UEGF, BMP1, LNK linker, MG macroglobulin, TED thioester-containing domain, C345c C-terminal domain. Peptides 75–85, 741–748, 905–913 locate at MG1, LNK, and MG7, respectively, are shown as spheres. Reposition of the TED (blue spheres in C3; green in C3b) shields residues 75–85, transitioning from an exposed state in C3 ( c ; red spheres) to a buried state in C3b ( d ; yellow spheres). Removal of C3a (cyan spheres) and relocation of TED (blue spheres in C3; green in C3b) shifts region of 741–748 from buried state at C3 ( e ; purple spheres), to an exposed conformation at C3b ( f ; yellow spheres). Conformational change of peptide 905–913 from C3 ( g ; purple spheres) to C3b ( h ; yellow spheres), due to reposition of CUB (blue spheres in C3; green in C3b). i Complement activation in T2DM was confirmed by ELISA ( n = 8, biological replicates). In the violin plot, the central dashed line indicates the median, and the upper and lower dashed lines indicate the quartiles. j Fold change (T2DM VS. WT) of C3 and C3b level ( n = 8, biological replicates). Error bars represent mean ± standard deviation (SD) across replicates. Structures of native C3 (PDB ID: 2A73) and C3b (PDB ID: 2I07). For all figures, * P < 0.05, ** P < 0.01, *** P < 0.001 by two tailed unpaired T -test with Welch’s correction. Exact P values are available in Supplementary Tables – . T2DM type 2 diabetes mellitus, WT wild type. Source data are provided as a Source Data file.

Article Snippet: To assess quantitatively the amount of native complement C3 and its cleaved and activated fragments (C3b), and thus the activation of the complement cascade, plasma of 8 male mice of each condition (WT and T2DM) were sampled in EDTA tubes and kept on ice to avoid ex vivo activation, C3 and C3b were measure with C3 and C3b mouse ELISA Kit (Prod# HK2002 for C3 and HK216 for C3b, Hycult, Uden, The Netherlands) respectively , , following the manufacturer’s instructions.

Techniques: Activation Assay, Enzyme-linked Immunosorbent Assay, Standard Deviation, Two Tailed Test