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hybridization and wash buffers  (LGC Biosearch)

 
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    LGC Biosearch hybridization and wash buffers
    Hybridization And Wash Buffers, supplied by LGC Biosearch, 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/hybridization+buffer/hybridization+buffer/pm28128342-275-5-10
    Average 90 stars, based on 1 article reviews
    hybridization and wash buffers - by Bioz Stars, 2026-11
    90/100 stars

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    Related Articles

    Incubation:

    Article Title: Retrotransposons hijack alt-EJ for DNA replication and eccDNA biogenesis.
    Article Snippet: Retrotransposons are highly enriched in the animal genome.. The activation of retrotransposons can rewrite host DNA information and fundamentally impact host biology.. Although developmental activation of retrotransposons can offer benefits for the host, such as against virus infection, uncontrolled activation promotes disease or potentially drives ageing.

    Article Title: Nanoscale cellular organization of viral RNA and proteins in SARS-CoV-2 replication organelles
    Article Snippet: For staining without CellMask (Thermo Fisher, C10046), cells were permeabilized in 70% ethanol at 4°C overnight. .. For CellMask staining, cells were permeabilized in 0.1% Triton X-100 at room temperature for 30 min. For RNA FISH staining, permeabilized cells were washed with 200 μL Wash Buffer A [40 μL Stellaris RNA FISH Wash Buffer A (LGC Biosearch Technologies, SMF-WA1-60), 20 μL deionized formamide, 140 μL H2O] at room temperature for 5 min, and incubated with 110 μL Hybridization Buffer [99 μL Stellaris RNA FISH Hybridization Buffer (LGC Biosearch Technologies, SMF-HB1-10), 11 μL deionized formamide] containing 1.1 μL 12.5 μM vgRNA FISH probes for 4 hours at 37°C in the dark. .. Then cells were washed with Wash Buffer A for 30 min at 37°C in the dark, washed with Wash Buffer A containing DAPI for 30 min at 37°C in the dark, and stored in Wash Buffer B (LGC Biosearch Technologies, SMF-WB1-20) for imaging.

    Article Title: The lncRNA Sweetheart regulates compensatory cardiac hypertrophy after myocardial injury in murine males
    Article Snippet: Prior to hybridization the coverslips were washed with diluted wash buffer A (LGC Biosearch Technologies #SMF-WA1–60) containing 10% deionized formamide (Merck #F9037). .. The coverslips were immersed in a hybridization buffer (LGC Biosearch Technologies #SMF-HB1-10) containing 10% deionized formamide and 1:100 smRNA FISH probe (Stellaris) and incubated at 37 °C o/n. .. After washing the coverslips at 37 °C for 30 min in the dark, they were washed again adding Hoechst DNA stain (Thermo Fisher Scientific #H3570) at 37 °C for 30 min in the dark.

    Hybridization:

    Article Title: Retrotransposons hijack alt-EJ for DNA replication and eccDNA biogenesis.
    Article Snippet: Retrotransposons are highly enriched in the animal genome.. The activation of retrotransposons can rewrite host DNA information and fundamentally impact host biology.. Although developmental activation of retrotransposons can offer benefits for the host, such as against virus infection, uncontrolled activation promotes disease or potentially drives ageing.

    Article Title: Nanoscale cellular organization of viral RNA and proteins in SARS-CoV-2 replication organelles
    Article Snippet: For staining without CellMask (Thermo Fisher, C10046), cells were permeabilized in 70% ethanol at 4°C overnight. .. For CellMask staining, cells were permeabilized in 0.1% Triton X-100 at room temperature for 30 min. For RNA FISH staining, permeabilized cells were washed with 200 μL Wash Buffer A [40 μL Stellaris RNA FISH Wash Buffer A (LGC Biosearch Technologies, SMF-WA1-60), 20 μL deionized formamide, 140 μL H2O] at room temperature for 5 min, and incubated with 110 μL Hybridization Buffer [99 μL Stellaris RNA FISH Hybridization Buffer (LGC Biosearch Technologies, SMF-HB1-10), 11 μL deionized formamide] containing 1.1 μL 12.5 μM vgRNA FISH probes for 4 hours at 37°C in the dark. .. Then cells were washed with Wash Buffer A for 30 min at 37°C in the dark, washed with Wash Buffer A containing DAPI for 30 min at 37°C in the dark, and stored in Wash Buffer B (LGC Biosearch Technologies, SMF-WB1-20) for imaging.

    Article Title: EGFR-induced lncRNA TRIDENT promotes drug resistance in non-small cell lung cancer via phospho-TRIM28-mediated DNA damage repair.
    Article Snippet: Coverslips were washed two times with PBS, fixed in 3.7% formaldehyde in PBS for 10 min at RT, followed by washing two times with PBS and immersed in 70% EtOH at 4 °C for a minimum of 1 h. Prior hybridization, coverslips were washed with 2 mL of wash buffer A (LGC Biosearch Technologies) supplemented with 10% deionized formamide (Invitrogen) at RT for 5 min. .. Cells were hybridized with 80 μL of hybridization buffer (LGC Biosearch Technologies) supplemented with 10% deionized formamide containing 1:100 dilution of smRNA FISH probes overnight at 37 °C in a humid chamber in the dark. ..

    Article Title: The lncRNA Sweetheart regulates compensatory cardiac hypertrophy after myocardial injury in murine males
    Article Snippet: Prior to hybridization the coverslips were washed with diluted wash buffer A (LGC Biosearch Technologies #SMF-WA1–60) containing 10% deionized formamide (Merck #F9037). .. The coverslips were immersed in a hybridization buffer (LGC Biosearch Technologies #SMF-HB1-10) containing 10% deionized formamide and 1:100 smRNA FISH probe (Stellaris) and incubated at 37 °C o/n. .. After washing the coverslips at 37 °C for 30 min in the dark, they were washed again adding Hoechst DNA stain (Thermo Fisher Scientific #H3570) at 37 °C for 30 min in the dark.

    Article Title: VarID2 quantifies gene expression noise dynamics and unveils functional heterogeneity of ageing hematopoietic stem cells
    Article Snippet: Cell pellet was resuspended in 200 μl of 70% ethanol, incubated at 4 °C for 1 h and then washed with 200 μl of wash buffer A (LGC Biosearch Technologies, SMF-WA1-60) supplemented with 10% deionized formamide (Thermo Fisher Scientific, 4,440,753) at room temperature for 5 min. .. Cells were hybridized with 80 μl of hybridization buffer (LGC Biosearch Technologies, SMF-HB1-10) supplemented with 10% deionized formamide containing the FISH probes at a 1:100 dilution at 37 °C overnight. .. Cell pellet was resuspended in 200 μl of 70% ethanol, incubated at 4 °C for 1 h and then washed with 200 μl of wash buffer A (LGC Biosearch Technologies, SMF-WA1-60) supplemented with 10% deionized formamide (Thermo Fisher Scientific, 4,440,753) at room temperature for 5 min.

    Article Title: EDC-3 and EDC-4 Regulate Embryonic mRNA Clearance and Biomolecular Condensate Specialization
    Article Snippet: .. Fixed embryos were washed in 1 mL Stellaris Wash Buffer A (LGC Biosearch SMF-WA1-60) with 10% formamide (Sigma F9037) for 5 minutes and hybridized with 100 μL of Stellaris Hybridization Buffer (LGC Biosearch SMF-HB1-10) containing 22.3 nM smFISH probes (48 probes targeting cgh-1 coding sequence conjugated with Quasar 670 dye, see Table S2 , smFISH Probes tab) for ∼16 hours in the dark with shaking at 37°C. ..

    Article Title: The temporal dynamics of lncRNA Firre -mediated epigenetic and transcriptional regulation
    Article Snippet: Cover glasses were incubated in 70% ethanol at 4 °C for at least 1 h and then washed with 1 mL of wash buffer A (LGC Biosearch Technologies) at room temperature for 5 min. .. Cells were hybridized with 100 μL of hybridization buffer (LGC Biosearch Technologies) containing the FISH probes at a 1:100 dilution in a humid chamber at 37 °C overnight up to 16 h. The next day, cells were washed with 1 mL of wash buffer A at 37 °C for 30 min and stained with wash buffer A containing 10 μg/mL Hoechst 33342 (Thermo Fisher) at 37 °C for 30 min. Cover glasses were washed with 1 mL of wash buffer B (LGC Biosearch Technologies) at room temperature for 5 min, mounted with ProLong Gold (Thermo Fisher) on a glass slide, and left to curate at 4 °C overnight before proceeding to image acquisition. .. Image acquisition was performed using a DeltaVision Elite widefield microscope with an Olympus UPlanSApo 100 × /1.40-numerical aperture oil objective lens and a PCO Edge sCMOS camera.

    Staining:

    Article Title: Nanoscale cellular organization of viral RNA and proteins in SARS-CoV-2 replication organelles
    Article Snippet: For staining without CellMask (Thermo Fisher, C10046), cells were permeabilized in 70% ethanol at 4°C overnight. .. For CellMask staining, cells were permeabilized in 0.1% Triton X-100 at room temperature for 30 min. For RNA FISH staining, permeabilized cells were washed with 200 μL Wash Buffer A [40 μL Stellaris RNA FISH Wash Buffer A (LGC Biosearch Technologies, SMF-WA1-60), 20 μL deionized formamide, 140 μL H2O] at room temperature for 5 min, and incubated with 110 μL Hybridization Buffer [99 μL Stellaris RNA FISH Hybridization Buffer (LGC Biosearch Technologies, SMF-HB1-10), 11 μL deionized formamide] containing 1.1 μL 12.5 μM vgRNA FISH probes for 4 hours at 37°C in the dark. .. Then cells were washed with Wash Buffer A for 30 min at 37°C in the dark, washed with Wash Buffer A containing DAPI for 30 min at 37°C in the dark, and stored in Wash Buffer B (LGC Biosearch Technologies, SMF-WB1-20) for imaging.

    Article Title: The temporal dynamics of lncRNA Firre -mediated epigenetic and transcriptional regulation
    Article Snippet: Cover glasses were incubated in 70% ethanol at 4 °C for at least 1 h and then washed with 1 mL of wash buffer A (LGC Biosearch Technologies) at room temperature for 5 min. .. Cells were hybridized with 100 μL of hybridization buffer (LGC Biosearch Technologies) containing the FISH probes at a 1:100 dilution in a humid chamber at 37 °C overnight up to 16 h. The next day, cells were washed with 1 mL of wash buffer A at 37 °C for 30 min and stained with wash buffer A containing 10 μg/mL Hoechst 33342 (Thermo Fisher) at 37 °C for 30 min. Cover glasses were washed with 1 mL of wash buffer B (LGC Biosearch Technologies) at room temperature for 5 min, mounted with ProLong Gold (Thermo Fisher) on a glass slide, and left to curate at 4 °C overnight before proceeding to image acquisition. .. Image acquisition was performed using a DeltaVision Elite widefield microscope with an Olympus UPlanSApo 100 × /1.40-numerical aperture oil objective lens and a PCO Edge sCMOS camera.

    Fluorescence In Situ Hybridization:

    Article Title: Nanoscale cellular organization of viral RNA and proteins in SARS-CoV-2 replication organelles
    Article Snippet: For staining without CellMask (Thermo Fisher, C10046), cells were permeabilized in 70% ethanol at 4°C overnight. .. For CellMask staining, cells were permeabilized in 0.1% Triton X-100 at room temperature for 30 min. For RNA FISH staining, permeabilized cells were washed with 200 μL Wash Buffer A [40 μL Stellaris RNA FISH Wash Buffer A (LGC Biosearch Technologies, SMF-WA1-60), 20 μL deionized formamide, 140 μL H2O] at room temperature for 5 min, and incubated with 110 μL Hybridization Buffer [99 μL Stellaris RNA FISH Hybridization Buffer (LGC Biosearch Technologies, SMF-HB1-10), 11 μL deionized formamide] containing 1.1 μL 12.5 μM vgRNA FISH probes for 4 hours at 37°C in the dark. .. Then cells were washed with Wash Buffer A for 30 min at 37°C in the dark, washed with Wash Buffer A containing DAPI for 30 min at 37°C in the dark, and stored in Wash Buffer B (LGC Biosearch Technologies, SMF-WB1-20) for imaging.

    Article Title: EGFR-induced lncRNA TRIDENT promotes drug resistance in non-small cell lung cancer via phospho-TRIM28-mediated DNA damage repair.
    Article Snippet: Coverslips were washed two times with PBS, fixed in 3.7% formaldehyde in PBS for 10 min at RT, followed by washing two times with PBS and immersed in 70% EtOH at 4 °C for a minimum of 1 h. Prior hybridization, coverslips were washed with 2 mL of wash buffer A (LGC Biosearch Technologies) supplemented with 10% deionized formamide (Invitrogen) at RT for 5 min. .. Cells were hybridized with 80 μL of hybridization buffer (LGC Biosearch Technologies) supplemented with 10% deionized formamide containing 1:100 dilution of smRNA FISH probes overnight at 37 °C in a humid chamber in the dark. ..

    Article Title: The lncRNA Sweetheart regulates compensatory cardiac hypertrophy after myocardial injury in murine males
    Article Snippet: Prior to hybridization the coverslips were washed with diluted wash buffer A (LGC Biosearch Technologies #SMF-WA1–60) containing 10% deionized formamide (Merck #F9037). .. The coverslips were immersed in a hybridization buffer (LGC Biosearch Technologies #SMF-HB1-10) containing 10% deionized formamide and 1:100 smRNA FISH probe (Stellaris) and incubated at 37 °C o/n. .. After washing the coverslips at 37 °C for 30 min in the dark, they were washed again adding Hoechst DNA stain (Thermo Fisher Scientific #H3570) at 37 °C for 30 min in the dark.

    Article Title: VarID2 quantifies gene expression noise dynamics and unveils functional heterogeneity of ageing hematopoietic stem cells
    Article Snippet: Cell pellet was resuspended in 200 μl of 70% ethanol, incubated at 4 °C for 1 h and then washed with 200 μl of wash buffer A (LGC Biosearch Technologies, SMF-WA1-60) supplemented with 10% deionized formamide (Thermo Fisher Scientific, 4,440,753) at room temperature for 5 min. .. Cells were hybridized with 80 μl of hybridization buffer (LGC Biosearch Technologies, SMF-HB1-10) supplemented with 10% deionized formamide containing the FISH probes at a 1:100 dilution at 37 °C overnight. .. Cell pellet was resuspended in 200 μl of 70% ethanol, incubated at 4 °C for 1 h and then washed with 200 μl of wash buffer A (LGC Biosearch Technologies, SMF-WA1-60) supplemented with 10% deionized formamide (Thermo Fisher Scientific, 4,440,753) at room temperature for 5 min.

    Article Title: The temporal dynamics of lncRNA Firre -mediated epigenetic and transcriptional regulation
    Article Snippet: Cover glasses were incubated in 70% ethanol at 4 °C for at least 1 h and then washed with 1 mL of wash buffer A (LGC Biosearch Technologies) at room temperature for 5 min. .. Cells were hybridized with 100 μL of hybridization buffer (LGC Biosearch Technologies) containing the FISH probes at a 1:100 dilution in a humid chamber at 37 °C overnight up to 16 h. The next day, cells were washed with 1 mL of wash buffer A at 37 °C for 30 min and stained with wash buffer A containing 10 μg/mL Hoechst 33342 (Thermo Fisher) at 37 °C for 30 min. Cover glasses were washed with 1 mL of wash buffer B (LGC Biosearch Technologies) at room temperature for 5 min, mounted with ProLong Gold (Thermo Fisher) on a glass slide, and left to curate at 4 °C overnight before proceeding to image acquisition. .. Image acquisition was performed using a DeltaVision Elite widefield microscope with an Olympus UPlanSApo 100 × /1.40-numerical aperture oil objective lens and a PCO Edge sCMOS camera.

    other:

    Article Title: EDC-3 and EDC-4 regulate embryonic mRNA clearance and biomolecular condensate specialization.
    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit polyclonal anti-EDC-4 This paper (Capralogics) Rabbit #6753 production bleed Rabbit polyclonal anti-EDC-3 This paper (Capralogics) Rabbit #7503 production bleed Rabbit polyclonal anti-CGH-1 This paper (Capralogics) Rabbit #7199 production bleed Rabbit monoclonal anti-HA (C29F4) Cell Signaling Technology Cat#3700; RRID: AB_2242334 Mouse monoclonal anti-FLAG M2 Sigma Cat#F1804; RRID: AB_262044 Mouse monoclonal anti-V5 Invitrogen Cat#R960-25; RRID:AB_2556564 Mouse monoclonal anti-beta-Actin (8H10D10) Cell Signaling Technology Cat#3700; RRID: AB_2242334 Mouse monoclonal anti-alpha-Tubulin (DM1A) Abcam Cat#ab7291; RRID: AB_2241126 Mouse monoclonal anti-HA (12CA5) Goodman Cancer Center Hybridoma Core Facility N/A Mouse monoclonal anti-Myc (9E10) Goodman Cancer Center Hybridoma Core Facility N/A IRDye 800CW (Goat anti-Rabbit) LI-COR Cat#926–32211; RRID: AB_621843 IRDye 680RD (Goat anti-Mouse) LI-COR Cat#926–68070; RRID: AB_10956588 Fluorescent TrueBlot : Anti-Mouse IgDyLightTM 680 (Rat anti-Mouse) Rockland Cat#18-4417-32; RRID: AB_2610840 Bacterial and virus strains DH5a Competent Cells Thermo Fisher Scientific Cat#18265017 E. coli OP50 Shared Fermentation Facility, Pennsylvania State University N/A E.coli HT115 RNAi clones Kamath et al.106 N/A BL-21(DE3) Competent Cells Promega Cat#L1196 Chemicals, peptides, and recombinant proteins Agarose BioShop Cat#AGA001.1 Stellaris Wash Buffer A LGC Biosearch Cat#SMF-WA1-60 Stellaris Hybridization Buffer LGC Biosearch Cat#SMF-HB1-10 Stellaris Wash Buffer B LGC Biosearch Cat #SMF-WB1-20 Vectashield Vector Labs Cat#H-1000-10 Anti-FLAG M2 magnetic beads Sigma Cat#M8823 Anti-HA C29F4 magnetic beads Cell Signaling Technology Cat#3724S Anti-Myc 9E10 magnetic beads Thermo Scientific Cat#88842 Critical commercial assays TruSeq Stranded Total RNA Library Prep Gold Illumina Cat#20020599 iScriptTM Reverse Transcription Supermix Biorad Cat#1708841 SsoAdvanced Universal SYBRGreen Supermix Biorad Cat#1725274 Deposited data DCAP-2 LC-MS/MS on WT and edc-3/4 mutants This paper MassIVE: MSV000093990 RNA-seq on WT and edc-3/4 mutants This paper GEO: GSE273136 Raw data This paper Mendeley, https://doi.org/10.17632/ 6jzy7ynx95.1 Experimental models: Organisms/strains C. elegans strain: edc-3(ok1427), outcrossed 6x Caenorhabditis Genetics Center RB1311 (Continued on next page) 18 Cell Reports 43, 114781, October 22, 2024

    Sequencing:

    Article Title: EDC-3 and EDC-4 Regulate Embryonic mRNA Clearance and Biomolecular Condensate Specialization
    Article Snippet: .. Fixed embryos were washed in 1 mL Stellaris Wash Buffer A (LGC Biosearch SMF-WA1-60) with 10% formamide (Sigma F9037) for 5 minutes and hybridized with 100 μL of Stellaris Hybridization Buffer (LGC Biosearch SMF-HB1-10) containing 22.3 nM smFISH probes (48 probes targeting cgh-1 coding sequence conjugated with Quasar 670 dye, see Table S2 , smFISH Probes tab) for ∼16 hours in the dark with shaking at 37°C. ..



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    Image Search Results


    (A, B) The C5-hFc-siSOD1 conjugate was dosed in WT mice as a single IV or SC bolus at 1.5 mg/kg (A) or 0.14 to 9 mg/kg siRNA equivalent dose (B). Target knockdown was measured 14 days after administration, using RT-qPCR on total RNA extracts from indicated regions of the CNS. The results are expressed as a ratio to mRNA levels measured in PBS-treated animals after normalization to RpL13 and RpL30 reference genes. (A) Box plots: extends from the 25th to 75th percentiles with center line plotted at the median. Whiskers indicate the smallest and the largest experimental values (n = 8-16 animals per group from 3 independent experiments). (B) Dose-response curves in each CNS region, where experimental results are expressed as means ± SEM, and estimated ED 50 values. (C) Mice were administered with either multi-IV (Q2Dx3) at 1.5 mg/kg or single SC at 4.5 mg/kg siRNA equivalent dose of the C5-hFc-siSOD1 conjugate. Shown are in situ hybridization micrographs showing siSOD1 AS distribution (top panels, miRNAScope®) throughout the brain (sagittal sections) and spinal cord (cross-sections), and corresponding Sod1 mRNA (lower panels, RNAScope®), 14 days after the last IV or single SC (representative animal from n=4 mice per group). (D) Magnifications and neuron-specific co-staining, using a Map2 mRNA-specific probe, in indicated regions of the brain.

    Journal: bioRxiv

    Article Title: Harnessing TfR1 for Cross-Species Systemic Delivery of siRNAs to Deep Brain Regions Using Single-Domain Antibodies

    doi: 10.64898/2026.05.20.726486

    Figure Lengend Snippet: (A, B) The C5-hFc-siSOD1 conjugate was dosed in WT mice as a single IV or SC bolus at 1.5 mg/kg (A) or 0.14 to 9 mg/kg siRNA equivalent dose (B). Target knockdown was measured 14 days after administration, using RT-qPCR on total RNA extracts from indicated regions of the CNS. The results are expressed as a ratio to mRNA levels measured in PBS-treated animals after normalization to RpL13 and RpL30 reference genes. (A) Box plots: extends from the 25th to 75th percentiles with center line plotted at the median. Whiskers indicate the smallest and the largest experimental values (n = 8-16 animals per group from 3 independent experiments). (B) Dose-response curves in each CNS region, where experimental results are expressed as means ± SEM, and estimated ED 50 values. (C) Mice were administered with either multi-IV (Q2Dx3) at 1.5 mg/kg or single SC at 4.5 mg/kg siRNA equivalent dose of the C5-hFc-siSOD1 conjugate. Shown are in situ hybridization micrographs showing siSOD1 AS distribution (top panels, miRNAScope®) throughout the brain (sagittal sections) and spinal cord (cross-sections), and corresponding Sod1 mRNA (lower panels, RNAScope®), 14 days after the last IV or single SC (representative animal from n=4 mice per group). (D) Magnifications and neuron-specific co-staining, using a Map2 mRNA-specific probe, in indicated regions of the brain.

    Article Snippet: For knock-down analysis on tissue sections using in situ hybridization (RNAScope, ACD), left cerebral hemisphere half-slices were cut into one ventral and one dorsal parts and immediately immersed in 10% neutral buffer formalin (NBF) fixative solution for approximately 48h at RT.

    Techniques: Knockdown, Quantitative RT-PCR, In Situ Hybridization, RNAscope, Staining

    (A-B) In situ hybridization micrographs showing siSOD1 AS distribution (miRNAScope®) throughout the brain (sagittal sections) and spinal cord (cross-sections) (A), 14 days after multi-IV bolus (Q2D x3) of the non-binding C5neg- or the TfR1-binding B8V32-hFc-siSOD1 conjugates, at 1.5 mg/kg siRNA. (B) In situ hybridization micrographs showing siSOD1 AS distribution (top panels, miRNAScope®) and corresponding Sod1 mRNA (lower panels, RNAScope®) in retina layers of the same animals (representative animal from n=4 mice per group). Retina layers: GCL, Ganglion Cell Layer; IPL, Inner Plexiform Layer; INL, Inner Nuclear Layer; OPL, Outer Plexiform Layer; ONL, Outer Nuclear Layer; IS, Inner Segments; OS, Outer Segments; RPE, Retinal Pigment Epithelium; Chor, Choroid.

    Journal: bioRxiv

    Article Title: Harnessing TfR1 for Cross-Species Systemic Delivery of siRNAs to Deep Brain Regions Using Single-Domain Antibodies

    doi: 10.64898/2026.05.20.726486

    Figure Lengend Snippet: (A-B) In situ hybridization micrographs showing siSOD1 AS distribution (miRNAScope®) throughout the brain (sagittal sections) and spinal cord (cross-sections) (A), 14 days after multi-IV bolus (Q2D x3) of the non-binding C5neg- or the TfR1-binding B8V32-hFc-siSOD1 conjugates, at 1.5 mg/kg siRNA. (B) In situ hybridization micrographs showing siSOD1 AS distribution (top panels, miRNAScope®) and corresponding Sod1 mRNA (lower panels, RNAScope®) in retina layers of the same animals (representative animal from n=4 mice per group). Retina layers: GCL, Ganglion Cell Layer; IPL, Inner Plexiform Layer; INL, Inner Nuclear Layer; OPL, Outer Plexiform Layer; ONL, Outer Nuclear Layer; IS, Inner Segments; OS, Outer Segments; RPE, Retinal Pigment Epithelium; Chor, Choroid.

    Article Snippet: For knock-down analysis on tissue sections using in situ hybridization (RNAScope, ACD), left cerebral hemisphere half-slices were cut into one ventral and one dorsal parts and immediately immersed in 10% neutral buffer formalin (NBF) fixative solution for approximately 48h at RT.

    Techniques: In Situ Hybridization, Binding Assay, RNAscope

    Cynomolgus monkeys were dosed by 30-min IV infusion three times at 7 days intervals with the B8V32-hFc-siHPRT conjugate, at the same dose previously evaluated in hTfR-KI mice of 1.5 mg/kg siRNA equivalent dose. Plasma samples were collected at indicated timepoints for PK analysis (A). Tissues were recovered at 14 days after last injection to assess target knockdown using RT-qPCR (B) and in situ hybridization in the brain using RNAScope® on one coronal section through the putamen (top panels: approximately -1 to -3 mm from the anterior commissure, based on Calabrese et al. 2015) and one through the hippocampus (lower panels: approximately -11 to -13 mm from the anterior commissure; star indicates an artefact) (C). (D) Neuroinflammation was assessed on brain tissue samples by measurement of mRNA levels of IbA1, GFAP and C3 as markers of microglial reactivity, astrocyte reactivity and complement activation, respectively.

    Journal: bioRxiv

    Article Title: Harnessing TfR1 for Cross-Species Systemic Delivery of siRNAs to Deep Brain Regions Using Single-Domain Antibodies

    doi: 10.64898/2026.05.20.726486

    Figure Lengend Snippet: Cynomolgus monkeys were dosed by 30-min IV infusion three times at 7 days intervals with the B8V32-hFc-siHPRT conjugate, at the same dose previously evaluated in hTfR-KI mice of 1.5 mg/kg siRNA equivalent dose. Plasma samples were collected at indicated timepoints for PK analysis (A). Tissues were recovered at 14 days after last injection to assess target knockdown using RT-qPCR (B) and in situ hybridization in the brain using RNAScope® on one coronal section through the putamen (top panels: approximately -1 to -3 mm from the anterior commissure, based on Calabrese et al. 2015) and one through the hippocampus (lower panels: approximately -11 to -13 mm from the anterior commissure; star indicates an artefact) (C). (D) Neuroinflammation was assessed on brain tissue samples by measurement of mRNA levels of IbA1, GFAP and C3 as markers of microglial reactivity, astrocyte reactivity and complement activation, respectively.

    Article Snippet: For knock-down analysis on tissue sections using in situ hybridization (RNAScope, ACD), left cerebral hemisphere half-slices were cut into one ventral and one dorsal parts and immediately immersed in 10% neutral buffer formalin (NBF) fixative solution for approximately 48h at RT.

    Techniques: Clinical Proteomics, Injection, Knockdown, Quantitative RT-PCR, In Situ Hybridization, RNAscope, Activation Assay