goat anti human a1at antibody hrp conjugate  (Bethyl)

Journal: Nucleic Acids Research

Article Title: RNA editing for the treatment of alpha-1 antitrypsin deficiency

doi: 10.1093/nar/gkaf1518

Figure Lengend Snippet: SERPINA1-994 decreases polymeric AAT accumulation in human iPSC-derived ZZ hepatocytes. ( A ) Images of cultured iPSCs after 14 days of treatment with the indicated AIMer. AAT polymer staining (green) and DAPI staining of nuclei (blue) are shown. The scale bars are 100 μm. ( B ) Quantification of polymeric AAT after 5, 10, or 14 days of treatment with increasing concentrations (0, 1.25, 2.5, 5.0, or 10 µM) of the indicated AIMer. Data are presented as mean ± SEM ( n = 6). White-adjusted two-way ANOVA with post-hoc comparisons to untreated allowing heteroscedasticity. ( C ) SERPINA1 RNA editing after 5, 10, or 14 days of treatment. Data are presented as mean ± SEM ( n = 4–6). White-adjusted two-way ANOVA with post-hoc comparisons to untreated allowing for heterscedasticity. ( D ) Quantification of nuclei per field after 5, 10, or 14 days. Data are presented as mean ± SEM ( n = 6). Two-way ANOVA with post-hoc comparisons to untreated. For all panels, P = 0.0 indicates that calculated P- values were below the limit of the program (2.2 × 10 −16 ). ns: nonsignificant.

Article Snippet: Fixed plates were immunostained with an antibody specific to the polymeric form of AAT (Hycult, Cat. No. HM2289) and quantified using a CellInsight CX7 high-content analysis platform (Thermo Fisher Scientific), and data were presented as mean fluorescence intensity per cell.

Techniques: Derivative Assay, Cell Culture, Polymer, Staining

Journal: Nature Communications

Article Title: Stereo-random oligonucleotides enable efficient recruitment of ADAR in vitro and in vivo

doi: 10.1038/s41467-025-64434-7

Figure Lengend Snippet: A Repetitive 2´ modification of large stretches of nucleosides of an ON targeting a GAPDH ORF site. B Partial 2´ modification, only at pyrimidine nucleosides. C Representative assessment of ON stability with an ON degradation assay in 100% FBS (Comprehensive, uncropped images are in Source data file). D Editing performance of partially stabilized ON in RPE, NHA and NHBE cells. GAPDH ORF target. Interferon was only applied with the v25.1 ON. E Gymnotic uptake of ON v117.28 (1 µM and 0.2 µM) in HeLa cells. GAPDH ORF target. F Repair of the SERPINA1 E342K mutation causing α-1-antitrypsin (A1AT) deficiency in a HeLa cell model stably overexpressing SERPINA1 E342K cDNA. Editing yields and normalized total A1AT levels in the supernatant of SERPINA1 E342K-piggyBac HeLa cells after treatment with partially modified ON designed to repair the SERPINA1 E342K mutation. The mock ON was the GAPDH ORF-targeting ON v117.19 (see Supplementary Table ). Data in A, B, D-F is shown as the mean ± s.d., N = 3 independent experiments. Experiments in A , B , E and F were done in HeLa cells. All editing experiments were performed with 50 nM ON, each datapoint represents a biological replicate.

Article Snippet: Goat anti-human A1AT antibody-HRP conjugate (Bethyl Laboratories, A80-122P) was applied at 0.5 μg/mL in PBS (100 μL) and incubated at room temperature for 1 h with gentle shaking (50 rpm).

Techniques: Modification, Degradation Assay, Mutagenesis, Stable Transfection

Journal: Nature Communications

Article Title: Stereo-random oligonucleotides enable efficient recruitment of ADAR in vitro and in vivo

doi: 10.1038/s41467-025-64434-7

Figure Lengend Snippet: A Treatment scheme of the PiZZ (SERPINA1 E342K) A1AT deficiency mouse model. Mice were treated with a single dose of 0–5 mg/kg LNP-encapsulated SERPINA1 E342K-targeting ON v117.82 (40 nt, 5’-29-1-10 symmetry, 5’-C A A) via tail vein injection. LNP formulation was based on the FDA-approved siRNA drug Onpattro. Mice were sacrificed 3 days post injection. B Sanger editing yields from hepatocytes (blue) and serum A1AT (human, wildtype) concentrations (yellow) clearly show a dose dependent correction at RNA and protein level. The considered therapeutic threshold of 11 µM serum A1AT (ref. ) is depicted as the dotted horizontal line. C Correlation of in vivo editing yields and resulting wildtype A1AT serum concentration. Data in B is shown as the mean ± s.d., N = 2–5 independent animals per condition. Figure 5A contains an icon created in BioRender (Stafforst, T. (2025), https://BioRender.com/zfhahgx .

Article Snippet: Goat anti-human A1AT antibody-HRP conjugate (Bethyl Laboratories, A80-122P) was applied at 0.5 μg/mL in PBS (100 μL) and incubated at room temperature for 1 h with gentle shaking (50 rpm).

Techniques: Injection, Formulation, In Vivo, Concentration Assay