anti human alp  (Cell Signaling Technology Inc)

Journal: iScience

Article Title: Robust effector memory features of human T-bet hi B cells induced by repeated mRNA vaccination

doi: 10.1016/j.isci.2026.114804

Figure Lengend Snippet: RBD-specific T-bet hi B cells induced by multiple mRNA vaccinations are robustly affinity-matured and can rapidly differentiate into antibody-secreting cells (ASCs) (A) Gene set enrichment analysis (GSEA) showing that RBD-specific T-bet hi B cells are significantly enriched in gene sets related to effector functions and activation, compared with RBD-specific switched memory B cells. Dotted lines indicate false discovery rate (FDR) of 0.05. (B) Gating strategy for fluorescence-activated cell sorting (FACS) of T-bet hi and classical memory B cells. (C) Representative ASC frequencies among sorted T-bet hi and classical memory B cells at 48 h after in-vitro polyclonal stimulation with IL-2 and R848. (D) Summary of ASC frequencies after stimulation of sorted T-bet hi and classical memory B cells, showing more robust differentiation potentials of T-bet hi B cells. (E) Representative data from IgG ELISPOT with sorted memory B cell subpopulations. (F) Summary of Wuhan-Hu-1 RBD-specific IgG SFUs per 1000 total IgG SFUs normalized by RBD-specific B cell frequency of each subset between classical and T-bet hi B cells. (G) Summary of RBD-specific IgG SFUs per 1000 total IgG SFUs among stimulated classical memory, unstimulated classical memory and T-bet hi B cells. (H) Surrogate RBD neutralization capacity of antibodies secreted by memory B cell subsets, in terms of binding inhibition rates. (I) Heavy chain somatic hypermutation frequency of RBD-specific memory B cells 14 days after the 2 nd or 3 rd vaccination, showing significant and comparable affinity maturation of T-bet hi B cell. (J) LIBRA-seq scores calculated from RBD-tetramer ADT increases in both subpopulations following additional vaccine doses. Permutation test for GSEA terms in A, Wilcoxon rank-sum test for D–4H and Mann-Whitney U test for I and 4J, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001; n.s., non-significant. Pooled data are represented as mean ± SD.

Article Snippet: ELISpot Flex: Human IgG (ALP) , MABTECH , Cat# 3850-2A.

Techniques: Activation Assay, Fluorescence, FACS, In Vitro, Enzyme-linked Immunospot, Neutralization, Binding Assay, Inhibition, MANN-WHITNEY

Journal: Nature Communications

Article Title: Tailored collagen binding of albumin-fused hyperactive coagulation factor IX dictates in vivo distribution and functional properties

doi: 10.1038/s41467-025-62955-9

Figure Lengend Snippet: a Illustration of the ELISA-based binding assay used to study binding between hFcRn or mFcRn and FIX-HSA variants at pH 5.5. Soluble truncated FcRn (light gray) was captured on IgG1 MST/HN (blue) coated in the well, before FIX-HSA (red/black) was added to the wells, and the HSA region of the fusion protein was detected by an ALP-conjugated anti-HSA antibody (dark gray). b , c Results from the ELISA-based hFcRn and mFcRn binding assays performed at pH 5.5. Data represent the mean ± SD of technical duplicates from one representative experiment. d Illustration of the SPR experiment used to study the interaction between FcRn (gray) and FIX-HSA (orange/black) performed by immobilizing FIX-HSA fusion proteins on a CM5 sensor chip before injecting a truncated form of soluble FcRn in a concentration gradient. Representative sensorgrams from SPR performed by injecting soluble truncated hFcRn ( e – h ) or mFcRn ( i – l ) over immobilized FIX-HSA variants at pH 5.5, showing one out of three independent experiments performed. The dotted lines show curves fitted by the 1:1 Langmuir binding model. m An illustration of the HERA setup designed to study hFcRn-mediated cellular ( n ), uptake and ( o ), recycling of FIX-HSA fusion proteins. Data are presented as relative to FIX-HSA WT (=1, dotted line) and represent the group mean ± SD of three independent experiments performed in technical triplicates (uptake: n = 9, except n = 8 for FIX KA -HSA WT and FIX KR -HSA WT , n = 6 for Padua KR -HSA QMP ; recycling: n = 9, except n = 7 for FIX KA -HSA WT ). Statistical significance was tested by unpaired two-tailed Student’s t tests with 95% confidence level (precise p -values are given in Supplementary Table ). FIX-HSA WT , black; Padua-HSA QMP , gray; FIX KA -HSA WT , blue; FIX KR -HSA WT , orange ; Padua KA -HSA QMP , pink; Padua KR -HSA QMP , purple. Source data are provided as a Source Data file. a , d , m Created in BioRender. Hovden Aaen, K. (2025) https://BioRender.com/auh41ln .

Article Snippet: After subsequent incubation for 1 h at RT, the wells were washed as prior, before detection was performed by adding 125 ng/mL of an alkaline phosphatase (ALP)-conjugated goat polyclonal anti-HSA antibody (Bethyl Laboratories, Inc., #A80-229AP, lot 10) in PBSTM at 100 μL/well.

Techniques: Enzyme-linked Immunosorbent Assay, Binding Assay, Concentration Assay, Two Tailed Test